JP5698900B2 - Cell culture transfer device - Google Patents

Description

  The present invention relates to a device capable of easily performing a transplant operation of a cell culture, particularly a cell culture used for treatment of diseases and injuries in humans and animals, and the collection and delivery of the cell culture using the device. Relating to the method.

  Despite recent advances in the treatment of heart disease, a treatment system for severe heart failure has not yet been established. For the treatment of heart failure, medical treatment with β-blockers or ACE inhibitors is performed. For heart failure that has become so severe that these treatments are not successful, replacement therapy such as an artificial heart or a heart transplant, that is, surgical treatment Is done.

Severe heart failure that is the subject of such surgical treatment includes those caused by advanced valvular disease and severe myocardial ischemia, acute myocardial infarction and its complications, acute myocarditis, ischemic cardiomyopathy (ICM), dilation There are a wide variety of causes such as chronic heart failure due to dilated cardiomyopathy (DCM) and acute aversion.
Depending on these causes and severity, valvuloplasty and replacement, coronary artery bypass surgery, left ventricular plastic surgery, mechanical assisted circulation, etc. are applied.

  Of these, only heart transplantation or replacement treatment with an artificial heart has been regarded as an effective treatment for those who have suffered heart failure due to a severe decrease in left ventricular function caused by ICM or DCM. However, replacement therapy for these patients with severe heart failure has many problems to be solved, such as chronic donor shortages, the need for continuous immunosuppression, and the development of complications. Is hard to say.

  Due to this severe situation of heart transplantation, other surgical treatments such as Batista surgery have been tried for a while, and it has attracted much attention as an alternative treatment for heart transplantation. However, efforts have been made to improve surgical procedures and establish indications.

On the other hand, recently, development of a new regenerative treatment method is considered indispensable as a solution for the treatment of severe heart failure.
In severe myocardial infarction and the like, cardiomyocytes become dysfunctional, and fibroblast proliferation and interstitial fibrosis progress, resulting in heart failure. As the heart failure progresses, the cardiomyocytes are damaged and fall into apoptosis, but the cardiomyocytes hardly undergo cell division, so the number of cardiomyocytes decreases and the cardiac function further decreases.
Cell transplantation is considered useful for the recovery of cardiac function in such patients with severe heart failure, and clinical applications using autologous skeletal myoblasts have already begun in the United States and Europe.

  However, in order to actually improve the cardiac function sufficiently by the cell transplantation method, 70-80% of the transplanted cells are lost by the direct intramyocardial injection method, and the effect cannot be fully exhibited. For example, dilated cardiomyopathy because it is indispensable to solve problems such as side effects such as securing a large and safe cell source, as well as causing inflammation to the local site of cell injection and only local cell transplantation. There is a limit when the heart function of the whole heart is lowered.

  In recent years, in response to these problems, a three-dimensional cell culture that can be applied to the heart including cells derived from parts other than the adult myocardium by using a temperature-responsive culture dish applying tissue engineering, and The manufacturing method was provided (patent document 1).

  In order to transplant such a cell culture to the target affected area (transplant site), for example, the end of the cell culture is picked with tweezers, the cell culture is taken out of the packaging container, and transferred to the affected area. However, a series of operations such as transplantation (attachment) to the affected area is required. However, cell cultures have low absolute physical strength and are prone to wrinkling, tearing, and breakage. Requires advanced technology and requires careful attention.

In order to supplement the strength of cell cultures, supports using hydrophilic PVDF membranes, nitrocellulose membranes and supports based on human fibrinogen, etc. are also known. An instrument is provided, and a support (Cell Shifter ^™ , manufactured by Cellseed) for cell culture corresponding to the aforementioned temperature-responsive culture dish is commercially available.

For example, in Patent Document 2, a cultured cell moving jig having a cell attachment portion is used, and cultured cells on a cell culture substrate are placed on a cell attachment portion made of a cell adhesion protein, a cell adhesion polymer, a hydrophilic polymer, or the like. The cultured cells are detached from the cell culture substrate by attaching them, and then the detached cultured cells are moved to a specific location by weakening the adhesion between the cell attachment part of the cultured cell transfer jig and the cultured cells. Reattachment is described.
However, it is not easy to peel the cultured cells adhered to the cell adhesion protein (for example, fibrin) present on the surface of the cell adhesion part of the cultured cell migration jig. It is considered that the cultured cells are not detached from the cell adhesion protein unless the adhesion between the site to which the cultured cells are applied and the cultured cells is stronger. In Patent Document 2, in order to peel the cultured cells from the cell adhesion part of the cultured cell transfer jig, only the problem of weakening the adhesion between the cell adhesion part and the cultured cell is shown. The means to solve this problem is not specified. Therefore, this cell culture transfer jig has a problem in terms of operability because it is difficult to peel off the cultured cells adhered to the cell adhesion part.

Patent Document 3 describes a delivery device used when a cell culture is transplanted to a target affected area. This transport and administration device is a therapeutic device that performs a deformation operation into a flat deployment shape and a cylindrical storage shape, and a bending operation of the proximal end portion by a change in fluid pressure supplied to the inside, Specifically, when transplanting the cell culture into the affected area, the cell culture is held, transferred to the affected area, and attached to the affected area. However, this transport and administration device has a structure in which a fine fluid channel is provided inside the sheet support and the fluid channel must be strictly managed. There was a problem in terms.
In order to improve this problem, the delivery device of Patent Document 4 has been provided. The structure thereof includes a cylindrical outer cylinder, and a slide member supported in the outer cylinder so as to be slidable in the axial direction. A sheet-shaped therapeutic substance is supported to be provided at the tip of the slide member, and in a free state protruding from the tip of the outer cylinder, it is maintained in a flat developed shape, and is removed according to the sliding movement of the slide member. It has a sheet support member that is accommodated in the outer cylinder while being deformed into a cylindrical shape while abutting against the tip of the outer cylinder when moved in the inner direction of the cylinder, and is still complicated and easy to operate. It is hard to say that it is a transplantation device having sex.

JP 2007-528755 A JP 2005-176812 A JP 2008-173333 A JP 2009-511 A

An object of the present invention is to provide a transfer device having simple operability in place of a conventional support or transportation administration device in transplantation operation of a cell culture used for treatment of diseases and injuries in humans and animals. An object of the present invention is to provide a transfer device capable of transplanting (attaching) a cell culture easily, quickly and reliably with a simple configuration.

  The inventors of the present invention have made extensive studies in order to achieve the above-mentioned problems. As a result, the cell culture holding part having a hydrophobic cell culture holding surface having at least one opening, and a liquid through the opening are provided. A liquid suction part disposed on the opposite side of the cell culture holding surface of the cell culture holding part that can be aspirated, wherein the opening has the cell culture in the cell culture holding part. When the cell culture is brought into contact with the cell culture holding surface, an instrument configured to have at least one contact is used to collect the cell culture released in the liquid and to collect the target site. The present invention was completed. Accordingly, the present invention relates to the following.

(1) A device for collecting and delivering a cell culture released in a liquid, the cell culture holder having a hydrophobic cell culture holding surface having at least one opening; A liquid-absorbing part disposed on the opposite side of the cell culture holding surface of the cell culture holding part, wherein the liquid can be sucked through the opening, and the opening contains the cell culture The device, wherein the device is configured to have at least one contact with the cell culture holding surface when brought into contact with the cell culture holding portion.
(2) The device according to (1), further including a hollow housing, wherein the liquid absorbing portion is disposed inside the housing, and at least one outer surface of the housing is configured as a cell culture holding portion.
(3) The device according to (1) or (2) above, wherein the cell culture holding surface is more hydrophobic than the site for delivering the cell culture.
(4) The device according to any one of (1) to (3) above, wherein the cell culture holding surface has a porosity of 5 to 90% of the area.
(5) The instrument according to any one of (1) to (4) above, wherein the cell culture holding part has a form of a single porous body, a porous body, or a mesh.
(6) The instrument according to any one of (1) to (5) above, wherein the opening wall has a thickness of 0.1 to 2.0 mm.
(7) The instrument according to any one of (1) to (6), wherein the liquid absorbing part includes a liquid absorbing material.
(8) The device according to (7) above, wherein the liquid-absorbing material includes one or more materials selected from the group consisting of a porous body, a nonwoven fabric, a polymer material, a pulp material, and a paper material.
(9) The instrument according to any one of (1) to (8), further including an operation handle.
(10) The instrument according to any one of (7) to (9), wherein the housing is provided with a movable plate operable from the outside for draining the water-absorbing liquid-absorbing material.
(11) The instrument according to any one of (7) to (10), wherein the housing includes an opening / closing part for exchanging the liquid-absorbing material.
(12) A method for collecting and delivering a cell culture comprising:
(I) providing the device according to any one of (1) to (11) above,
(II) contacting the cell culture holding part of the device with the cell culture released in the liquid;
(III) a step of recovering the cell culture by adsorbing it to the cell culture holding unit together with the suction of the liquid by the liquid absorption unit;
(IV) moving the cell culture to a target site that is hydrophilic from the cell culture holding surface while adsorbing the cell culture to the cell culture holding unit;
Including said method.
(13) The method according to (12) above, wherein the target site has substantially no moisture on the surface thereof.

According to the present invention, by bringing the cell culture into contact with the holding part, the liquid present around the cell culture is sucked into the liquid absorbing part through the opening formed in the holding part, thereby The culture can be adsorbed to a hydrophobic holding surface. The adsorbed cell culture is held on the holding surface by the action of the surface tension of the absorbed liquid. In addition, the operation of bringing the cell culture held on the holding surface into contact with the target affected part (transplanted site) allows the liquid held in the liquid absorbing part to move to the affected part and transplant the cell culture to the target site. it can. Since the holding surface is hydrophobic, if the target transplant site is more hydrophilic than the holding surface, the cell culture can be easily detached from the holding surface and moved to the target site. In other words, if adhesion by surface tension between the cell culture and the target transplant site is superior to adhesion by surface tension between the cell culture and the hydrophobic holding surface, the cell culture is peeled off from the holding part to Can be transplanted to the transplant site. When excessive moisture is present at the intended transplantation site, an appropriate difference in surface tension can be formed by substantially removing the moisture.
Thus, according to the present invention, the cell culture can be transplanted (attached) to the affected area easily, quickly and reliably.
In addition, the instrument of the present invention has a simple configuration, can be sterilized (for example, ethylene oxide gas), and has very good operability.

FIG. 1 is a schematic side view of a device of the present invention and a schematic side view of a cell culture in an immersion liquid. FIG. 2 is a side cross-sectional view of the instrument of the present invention. FIG. 3 is a bottom view of the holding part of the instrument of the present invention. FIG. 4 is a top view of the device of the present invention. FIG. 5A is a view showing a draining operation of the water-absorbing liquid-absorbing material in the side sectional view of the housing portion of the instrument of the present invention. FIG. 5B is a view showing a drainage operation of the water-absorbing liquid-absorbing material in the side sectional view of the housing portion of the instrument of the present invention. The arrow indicates the movement of the movable plate. FIG. 6A is a front view of the instrument of the present invention showing the opening / closing operation when the liquid-absorbing material is replaced. FIG. 6B is a diagram showing an opening / closing operation at the time of exchanging the liquid-absorbing material in the front view of the instrument of the present invention. FIG. 7A is a side view showing a state of the cell culture in the immersion liquid. FIG. 7B is a side cross-sectional view of the instrument of the present invention, showing the state of adsorption of the cell culture using the instrument together with the cell culture in the immersion liquid. FIG. 7C is a side cross-sectional view of the instrument of the present invention showing a state of holding a cell culture using the instrument. FIG. 7D is a side cross-sectional view of the instrument of the present invention showing the state of cell culture exfoliation (transplantation) using the instrument. FIG. 8 is a bottom view of the holding portion showing an opening having a shape different from that of FIG. FIG. 9 is a bottom view (upper side) and a side view (lower side) of the device of the present invention having no housing, in which the liquid absorption part and the holding part are made of different members. FIG. 10 is a bottom view (upper side) and a side view (lower side) of the instrument of the present invention having no housing, in which the liquid absorption part and the holding part are made of the same member.

  The present invention relates to a device for collecting and delivering a cell culture released in a liquid, a cell culture holder having a hydrophobic cell culture holding surface having at least one opening; A liquid-absorbing part disposed on the opposite side of the cell culture holding surface of the cell culture holding part capable of sucking a liquid through the opening, wherein the opening contains the cell culture. The instrument is configured such that the cell culture has at least one contact with the cell culture holding surface when brought into contact with the cell culture holding unit.

  The cell culture in the present invention is typically composed only of cultured cells and their products, but supplements and / or supports a predetermined part of the living body (for example, an affected part) without departing from the object of the present invention. Various materials (complementary materials and support materials) can be included. The cell culture may be used for treatment of diseases and injuries in mammals such as humans and livestock. The cell culture is released in a liquid state, that is, in a culture solution (immersion solution) or a washing solution, and is not attached to the surface of the culture substrate. A cell culture may contain only one cell, but usually contains two or more cells. The shape of the cell culture is not particularly limited, and may be various shapes such as a sheet shape, a block shape, and a column shape. A cell culture usually has flexibility (softness), but is not limited thereto, and may be, for example, a slightly hard material. The cell culture may be cultured on a so-called temperature-responsive culture dish (for example, see JP-A-2-21865) and peeled off from the culture dish, or may be obtained by other methods. . The cell culture is derived from any cell type, such as, but not limited to, myoblasts (eg, skeletal myoblasts), cardiomyocytes, fibroblasts, synoviocytes, epithelial cells, endothelial cells, etc. Also good. In the present invention, a cell culture composed of, for example, skeletal myoblasts is preferable from the viewpoint of therapeutic utility and difficulty in handling by conventional techniques. The animal from which the cells constituting the cell culture are derived is also not particularly limited and includes, for example, humans, non-human primates, dogs, cats, pigs, horses, goats, sheep and the like.

The instrument of the present invention includes at least a cell culture holding part and a liquid absorption part.
The cell culture holding part is a part that adsorbs and holds the cell culture (a part that adsorbs and / or holds). When the cell culture contains a supplementary material or a support material, the cell culture is held by the retention between the cells contained therein and the product thereof and the cell culture holding part. The holding part has a structure that forms a boundary between the adsorbed and held cell culture and the liquid absorbing part, and may be a separate member or the same member as the liquid absorbing part. When it is a separate member, the holding part can have an arbitrary shape having a predetermined thickness. The holding portion may be, for example, a flat shape, a concave shape, a convex shape, or a mesh shape, and may be flexible or rigid. In a preferred aspect of the present invention, the holding portion may be configured as a part of a housing described later. Moreover, when it is the same member as a liquid absorption part, a holding | maintenance part can be constructed | assembled by forming the hole and groove | channel of predetermined depth in a part of liquid absorption part. Here, the thickness of the holding part or the depth of the hole or groove provided in the liquid absorbing part is preferably 0.1 to 2.0 mm, more preferably 0.1 to 1.0 mm. 1 aspect of this invention WHEREIN: It is preferable that the outer diameter of a holding | maintenance part is 1.0-10 cm, and it is more preferable that it is 2.0-6.0 cm.

  The cell culture holding part preferably has a hydrophobic cell culture holding surface on the side in contact with the cell culture. Due to the hydrophobicity of the holding surface, a difference in surface tension is likely to occur between the cell culture and the delivery site, and the cell culture adsorbed and held on the holding surface can be moved to the delivery site more easily. Is possible. In order to make the holding surface hydrophobic, for example, it is preferable to apply a fluororesin, which is a hydrophobic material having a low friction coefficient, as a constituent material of the cell culture holding part, or to coat the surface of the holding part. Examples of fluororesin include polytetrafluoroethylene, polychlorotrifluoroethylene, polyvinylidene fluoride, polyvinyl fluoride, perfluoroalkoxy fluororesin, tetrafluoroethylene / hexafluoropropylene copolymer, ethylene / tetrafluoroethylene Examples thereof include, but are not limited to, a copolymer and an ethylene / chlorotrifluoroethylene copolymer.

The holding surface may comprise at least one opening. The opening is not particularly limited in shape and size as long as the cell culture is configured to have at least one contact with the cell culture holding surface when the cell culture is brought into contact with the holding portion. That is, the openings are preferably shaped and sized so that the cell culture will always contact the holding surface somewhere. By comprising in this way, it becomes easy to move a cell culture from a holding part to a delivery site. Therefore, the opening may be constituted by, for example, a large number of small holes (see FIG. 3), a mesh mesh, or a slit-shaped gap having various shapes (see FIG. 8). . In addition, one or two or more members having a hydrophobic surface and a predetermined thickness are directly bonded to a liquid absorption part to be described later, and the assembly of the members or members is used as a cell culture holding part. The part where the object holding part does not exist and the liquid absorbing part is exposed may be an opening (see FIG. 9). Alternatively, when the cell culture holding part is the same member as the liquid absorption part as described above, holes or grooves formed in the liquid absorption part are openings (see FIG. 10). The opening ratio of the holding portion is typically 5 to 90% with respect to the surface area of the holding portion, but a suitable range can be selected in consideration of the hydrophobicity of the holding portion surface. The thickness of the opening wall, that is, the distance between the holding surface and the liquid absorbing part surface is preferably 0.1 to 2.0 mm, more preferably 0.1 to 1.0 mm.

The liquid absorbing part is arranged on the side of the cell culture holding part opposite to the cell culture holding surface. The shape, structure, material, etc. of the liquid absorption part are arbitrary as long as the liquid can be sucked through the opening. For example, the liquid absorption part may be constituted by a hollow member liquid-tightly connected to the cell culture holding, including a structure such as a syringe that can mechanically suck liquid. You may be comprised with a liquid material.
The liquid-absorbing material is not particularly limited, but a material capable of reversibly absorbing and discharging liquid is preferable. That is, a material that can absorb and drain liquid is preferable to a material that absorbs liquid but cannot drain. Examples of materials that can absorb and drain liquid include a liquid-absorbing and draining polymer. Specific examples include polyurethane, polyurethane foam, polyurethane sponge, polypropylene urethane sponge, nylon nonwoven fabric, melamine foam, cotton-like pulp, liquid absorbent paper, and the like, but are not limited to these examples.

  Moreover, a hydrophilic polymer is mentioned as a material which absorbs liquid but cannot drain. Examples of hydrophilic polymers include water-containing gels such as polyacrylamide, polydimethylacrylamide, polyacrylic acid and salts thereof, polyhydroxyethyl methacrylate, polyhydroxyethyl acrylate, polyvinyl alcohol, polyvinyl pyrrolidone, cellulose, carboxymethyl cellulose, and (meta ) Temperature-responsive polymers composed of homopolymers or copolymers containing acrylamide compounds, N-alkyl substituted (meth) acrylamide derivatives, N, N-dialkyl substituted (meth) acrylamide derivatives and / or vinyl ether derivatives. Therefore, in one embodiment of the present invention, the liquid-absorbing material is a homopolymer comprising a (meth) acrylamide compound, an N-alkyl substituted (meth) acrylamide derivative, an N, N-dialkyl substituted (meth) acrylamide derivative and / or a vinyl ether derivative. It does not contain temperature responsive polymers consisting of polymers or copolymers. In another embodiment of the present invention, the liquid-absorbing material may be polyacrylamide, polydimethylacrylamide, polyacrylic acid and salts thereof, polyhydroxyethyl methacrylate, polyhydroxyethyl acrylate, polyvinyl alcohol, polyvinyl pyrrolidone, cellulose, carboxymethyl cellulose, and the like. Contains no hydrogel. In yet another embodiment of the present invention, the liquid absorbing material does not include a hydrophilic polymer.

In one aspect of the present invention, the instrument further includes a hollow housing, the liquid absorbing portion is disposed inside the housing, and at least one outer surface of the housing is configured as a holding portion having at least one contact with the cell culture. May be.
In one embodiment of the present invention, the device of the present invention is preferably made of a sterilizable material. Sterilization techniques include, but are not limited to, gas sterilization with ethylene oxide gas, autoclave sterilization, boiling sterilization, and the like. In another aspect, the device of the present invention is provided in a sterilized state.
The device of the present invention may be disposable or reusable. In this case, the entire device of the present invention may be disposable or reusable, or only a part thereof, for example, the liquid absorbing part or the liquid absorbing material is disposable and the remaining part. May be reusable.
The device of the present invention can also be composed of a transparent or translucent material. Thereby, the operator of the instrument can visually recognize the position and state of the cell culture and the delivery site through the instrument, and can work without changing the line of sight and the posture.

The device of the present invention can also be used for the collection and delivery of materials other than cell cultures, such as artificial tissues that do not contain cellular components, such as artificial dura mater, artificial cornea, artificial skin. The device of the present invention has a small mechanical effect on the recovered / delivered material, and is therefore particularly suitable for recovering / delivering cell cultures and materials having a fragile structure.
In addition, the device of the present invention can be used not only for transplanting a cell culture used for treatment of human and animal diseases and injuries, but also for collecting and delivering various materials released in the liquid. Therefore, the instrument of the present invention is not limited to medical use, but can also be used for research use and industrial use.

The present invention also provides
(I) providing the instrument;
(II) contacting the cell culture holding part of the device with the cell culture released in the liquid;
(III) a step of recovering the cell culture by adsorbing it to the cell culture holding unit together with the suction of the liquid by the liquid absorption unit;
(IV) moving the cell culture to a target site that is hydrophilic from the cell culture holding surface while adsorbing the cell culture to the cell culture holding unit;
To a method for harvesting and delivering cell cultures.
In step (IV) above, the movement is preferably performed in air.

The method further comprises:
(V) contacting the cell culture with the target site, thereby detaching the cell culture from the holder;
(VI) transplanting the cell culture to the target surface;
May be included.
The method may further include (V ′) substantially removing moisture at the target site prior to step (V). Therefore, the target site after the same step has substantially no moisture on the surface.

Hereinafter, although the instrument of this aspect is demonstrated in detail based on the suitable embodiment example shown to an accompanying drawing, this invention is not limited to this example.
FIG. 1 is a schematic side view of a device A of the present invention, a schematic side view of a cell culture being immersed in a culture solution, FIG. 2 is a side sectional view of the device A of the present invention, and FIG. FIG. 4 is a top view of the device A of the present invention, and FIGS. 5A and 5B are side cross-sectional views of the housing portion of the device A of the present invention, showing the drainage operation of the water-absorbing liquid-absorbing material. 6A and 6B are schematic views showing an operation for opening and closing the liquid-absorbing material at the front portion of the device A of the present invention. 7A, B, C, and D are schematic side views of the device A of the present invention. Adsorption, retention, and exfoliation (transplantation) of the cell culture using the device A together with the cell culture immersed in the culture medium. It is the schematic which showed the mode of. FIG. 8 is a bottom view of the holding portion showing an opening having a shape different from that of FIG. FIG. 9 is a bottom view and a side view of the instrument of the present invention having no housing, in which the liquid absorption part and the holding part are made of different members. FIG. 10 is a bottom view and a side view of the instrument of the present invention having no housing, in which the liquid absorption part and the holding part are made of the same member.

  The instrument A shown in these figures is an instrument (apparatus) used when transplanting (attaching) the cell culture 4 to a living body such as a human or an animal, that is, an affected part (transplanted site) of the living body. . The instrument A typically includes a housing 1, a handle 2, and a drainage operation unit 3.

  The housing 1 has the drainage operation unit 3 on the upper surface 15, and a movable plate 31 that is interlocked with the drainage operation unit 3 is disposed in a hollow interior and includes the liquid absorbing material 11. The movable plate 31 interlocked with the drainage operation unit 3 can be moved along the groove 16 provided on the upper surface 15 in the direction of compressing the liquid absorbent material 11. Further, a drainage port 14 is provided on the front surface 17 and further has an opening / closing part 18 necessary for taking in and out the liquid absorbent material 11. A plurality of openings 13 are formed in the holding portion 12.

  Although the number of grooves provided on the upper surface 15 of the housing 1 is one in the drawing, the number is not limited to this, and a plurality of grooves may be provided.

  The shape and quantity of the drainage port 14 provided on the front surface 17 of the housing 1 are two rectangular ones in the drawing, but are not limited to this, and a single one of circular, elliptical, or polygonal ones, or It is good also as three or more. Further, the installation of the drain port 14 is not limited to the front surface 17 of the housing 1. Furthermore, the drainage port 14 is installed as necessary and is not necessarily required.

  The structure of the opening / closing part 18 on the front surface 17 of the housing 1 is not particularly limited, and examples thereof include screwing and uneven fitting.

  In addition, although the shape of the housing 1 has comprised the cube shape in the structure shown in the figure, it is not restricted to this, For example, you may make polygonal shape other than the said cube, and cylindrical shape. Moreover, all or a part of the housing 1 can be made of a flexible material and can be configured to be deformable. With such a configuration, access to the delivery site can be facilitated or facilitated, or the cell culture can be better fitted to the delivery site by being deformed according to the shape of the delivery site.

  The constituent material of the housing 1 is not particularly limited. For example, polycarbonate, polymethyl methacrylate, methyl methacrylate / styrene, propylene, acrylonitrile-butadiene-styrene copolymer, polystyrene, epoxy resin, polyarylate, polysulfone, polyethersulfur, and the like. Examples thereof include polymer materials such as phon, polyamide, polybutylene terephthalate, fluororesin, poly-4-methylpentene-1, phenoxy resin, polyimide resin, and organic nonlinear optical material.

  The surface 121 of the holding part 12 is preferably composed of a hydrophobic material. For example, it is preferable to apply a fluororesin, which is a hydrophobic material having a low friction coefficient, as a constituent material, or to coat the surface of the holding portion. For example, polytetrafluoroethylene, polychlorotrifluoroethylene, polyvinylidene fluoride, polyvinyl fluoride, perfluoroalkoxy fluororesin, ethylene tetrafluoride / hexafluoropropylene copolymer, ethylene / tetrafluoroethylene copolymer, ethylene -Although a chlorotrifluoroethylene copolymer is mention | raise | lifted, it is not limited to these illustrations. Thereby, peeling operation of the adsorbed and held cell culture 4 can be performed more easily. The holding part may be a flexible flat plate or mesh having one or more small holes. The opening ratio of the holding part is 5 to 90% with respect to the surface area of the holding part, but an appropriate range can be selected in consideration of the hydrophobicity of the surface of the holding part.

  The shape of the liquid-absorbing material 11 is a rectangular parallelepiped in the drawing, but is not limited thereto, and may be a cylindrical shape or a polygonal cylindrical shape other than the rectangular parallelepiped.

  The handle 2 has an inclination angle (an angle formed between the normal line 19 and the axis 21) θ (see FIG. 2) of the normal line 19 of the housing 1 with respect to the axis 21 of the handle 2 in consideration of the convenience of actual operation. It is preferably about 90 °, and more preferably about 20 to 40 °.

  The shape of the handle 2 is a rectangular parallelepiped in the drawing, but is not limited to this, and may be a cylindrical body, a spatula-shaped or a scoop-shaped atypical body, or a shape considering ergonomics.

  The constituent material of the handle 2 is not particularly limited. For example, a polymer material such as polycarbonate, polypropylene, polyethylene, polystyrene, urethane, acrylonitrile-butadiene-styrene copolymer, methyl methacrylate / butadiene / styrene, polymethyl methacrylate, and the like. And metal materials such as stainless steel, aluminum, and titanium.

  Further, the constituent material of the drainage operation unit 3 is not particularly limited. For example, polycarbonate, polypropylene, polyethylene, polystyrene, urethane, acrylonitrile-butadiene-styrene copolymer, methyl methacrylate / butadiene / styrene, polymethyl methacrylate And polymer materials such as stainless steel, aluminum and titanium.

  In another aspect of the present invention, the drainage operation unit 3 may be provided in a part other than the housing, for example, a handle. In this case, the drainage operation unit 3 may be connected to the movable plate 31 and a connecting member such as a rod, or pneumatically or hydraulically. Alternatively, the movable plate 31 may be electrically operated, and the drainage operation unit 3 may be configured as a switch or a lever for electrically controlling the movable plate 31. The handle has a structure consisting of at least two layers, one layer is connected to the movable plate, another layer is connected to the housing, and the layer connected to the movable plate can slide along the layer connected to the housing. It can also be set as a simple structure. This layer is concentric, for example, a combination of an outer cylinder and an inner cylinder slidably disposed therein, or a relatively wide member and a groove provided therein. The combination with the relatively narrow member arrange | positioned so that sliding is possible may be sufficient. In this case, it is easy to operate if the layer connected to the movable plate and the non-connected end of the layer connected to the housing are provided with a grip that can be put with a finger or pressed with a palm. The shape of the grip is not particularly limited, and for example, it can be a ring shape into which a finger can be inserted. Further, the movable plate (for example, the state shown in FIG. 5B) moved in the direction of drainage is caused by the elasticity of the water absorbing material itself, the action of an elastic member such as a spring, and / or the action of an electric member that moves the movable plate. It is also possible to adopt a configuration in which the position automatically returns to the original position (for example, the state of FIG. By providing the drainage operation part on the support part of the instrument of the present invention, for example, the handle, various operations including the drainage operation can be performed with one hand, which is extremely advantageous when used for surgery, for example. . In an embodiment that does not have a housing, for example, the drainage operation can be performed with one hand by directly grasping the liquid-absorbing material with a hand in the manner of draining the sponge water.

  In an embodiment without a housing, the handle can be connected to the cell culture holding part and / or the liquid absorbing part. In this case, the handle may be formed integrally therewith, or may be configured as a separate member and connected to each other by a conventional method. Further, a liquid-permeable support member that supports both of them may be provided between the cell culture holding part and the liquid absorption part, and a handle may be connected thereto. The connection angle of the handle is the same as described above.

  Next, an example of a usage method (action) of the instrument A will be described. Here, it is assumed that the affected part to which the cell culture 4 is transplanted is, for example, a myocardial infarction site, and the cell culture 4 to be transplanted is, for example, a cell culture for improving cardiac function. To do.

First, the operator prepares the instrument A, and positions both of the cells so that the center of the cell culture 4 is positioned at the center of the holding part 12 on the cell culture 4 immersed in the culture solution 5 in the culture vessel 6. The relationship is adjusted and the cell culture 4 is brought into contact with the holder 12. Thereby, the culture solution (immersion solution) 5 of the cell culture 4 is absorbed into the liquid absorbent material 11 through the opening 13 of the holding unit 12, and the cell culture 4 is adsorbed to the holding unit 12 accordingly. At this time, in the culture vessel 6, the culture solution 5 in which the cell culture 4 is immersed decreases to such an extent that the retention between the cell culture 4 and the holding unit 12 is not affected as a result of being sucked. ing.
In an embodiment that does not have the liquid-absorbing material 11, the liquid absorption can be performed by a liquid absorption mechanism such as a syringe.

  In order to cause the absorbent material 11 to absorb the immersion liquid 5 of the cell culture 4, the space occupancy of the liquid absorbent material 11 provided in the housing 1 is preferably 50 to 100%, and 70 to 95%. It is more preferable that Moreover, it is preferable that the thickness of the opening 13 of the holding | maintenance part 12 is 0.1-2.0 mm, and it is more preferable that it is 0.1-1.0 mm.

Next, the holding part 12 to which the cell culture 4 is adsorbed is pulled up into the air. By doing so, the cell culture 4 is held in the air while adsorbed to the holding portion 12 of the housing 1.
This is because the surface of the cell culture 4 adsorbed on the holding part 12 is in contact with air (gas), but the cell culture 4 contains the immersion liquid 5 (liquid), so that the gas is more liquid than the liquid. Since the molecular density is small, the cell culture 4 is subjected to surface tension that is pulled only from molecules inside the immersion liquid 5, not from air-side molecules. Thus, an excellent holding effect can be obtained by adsorbing the cell culture 4 together with the immersion liquid 5.

  Next, while maintaining the state in which the cell culture 4 is adsorbed to the holding unit 12, the cell culture 4 is transferred to the affected part 7 (transplant site). The transfer is preferably carried out in gas in order to maintain good adsorption.

  Here, the holding part 12 of the separately prepared instrument A (the instrument in a state before holding the cell culture 4) can be brought into contact with the surface of the affected part 7 to remove moisture such as body fluid covering the affected part 7 surface. The removal of moisture is not limited to the device A, but can be performed by other water-absorbing materials such as gauze, non-woven fabric, and paper waste without limitation. Immediately after the removal, the holding part 12 of the device A on which the cell culture 4 is first adsorbed is brought into contact with the affected part, and the cell culture 4 is pressed. Thereby, the immersion liquid 5 covering the surface of the cell culture 4 moves to the affected area 7, and the cell culture 4 is affixed to the affected area 7 accordingly.

  This is based on the physical principle that the immersion liquid 5 covering the surface of the cell culture 4 is transferred to the affected area 7 from which water has been removed, because the holding surface 121 is made of a hydrophobic material. The cell culture 4 can be simply moved to the affected area 7 and attached. Due to the hydrophobicity of the surface of the holding part, the adhesion due to the surface tension between the cell culture and the affected part is superior to the adhesion due to the surface tension between the holding part and the cell culture, so that the cell culture can be transplanted to the affected part. . At this time, the amount of the immersion liquid 5 that moves to the affected part 7 is only the part that covers the cell culture 4 on the outer surface of the holding part 12, and is absorbed into the liquid-absorbing material 11 provided inside the housing 1. Since there is almost no movement of the immersion liquid 5, there is no fear that the cell culture 4 attached to the affected area will be washed away by an excessive amount of the immersion liquid 5.

  In the operation of transplanting into the affected part 7, the height of the housing 1 is suppressed to 0.5 to 5.0 cm, and an appropriate angle is applied to the connecting part with the handle 2, so that the affected part 7 is, for example, on the side of the heart. Even if it exists, the housing 1 can be made to follow reliably. The height of the housing 1 is preferably 1.0 to 3.0 cm. Moreover, it is preferable that the holding | maintenance part outer diameter is 1.0-10 cm, and it is more preferable that it is 2.0-6.0 cm.

The immersion liquid 5 contained in the liquid-absorbing material 11 provided inside the housing 1 is moved by moving the drainage operation unit 3 disposed on the upper surface 15 of the housing 1 along the groove 16 provided on the upper surface 15. The movable plate 31 inside the housing 1 interlocked with the drainage operation unit 3 moves in the direction in which the liquid-absorbing material 11 is compressed, and can be drained from the drainage port 14 provided on the front surface 17 of the housing 1. . Thereby, it becomes possible to repeatedly perform a series of operations of adsorption, holding and pasting of the cell culture 4.
In an embodiment without a housing, the drainage operation can be performed, for example, by manually pressing the water absorption part.

  Furthermore, the housing 1 has an opening / closing part 18 through which the liquid absorbent material 11 can be taken in and out, and when the liquid absorbent material 11 is deteriorated or broken, it can be replaced quickly. Without preparing the device A, it is possible to continue the operation of adsorbing, holding, and sticking the cell culture 4 with a simple replacement operation.

Example 1
In order to confirm the effect of the device A, the following experiment was performed.
The holding portion 12 has a diameter φ4 cm, a height of 2 cm, a side surface thickness of 3.0 mm, an upper surface thickness of 3.0 mm, and a holding portion thickness of 1.0 mm in a hollow rectangular parallelepiped hydrophobic housing (Teflon®), On the other hand, holes with a diameter of 1.0 mm were evenly arranged so as to have an opening ratio of 20%. A polyurethane sponge was housed in the housing so that the space occupancy was 90%, and a device 1 was obtained.

Comparative Example 1
A commercially available cell culture support (CellShifter ^™ , manufactured by Cellseed Co., Ltd., diameter 3 cm) was prepared.

For Experimental Examples 1 and 2, a series of operations of adsorption, retention, and transplantation (attachment) was performed on a cell culture having a diameter of 2.7 to 3.1 cm and a thickness of 400 to 470 μm, respectively.

  In the instrument 1, it was possible to immediately adsorb and easily hold the holding part while maintaining its shape only by lightly contacting the holding part with the cell culture. Furthermore, after removing moisture at the transplantation site with a separately prepared instrument 1, the cell 1 is immediately pasted without losing its shape simply by bringing the holding part of the instrument 1 holding the cell culture into contact with the transplantation site. I was able to.

  On the other hand, in the instrument 2, the support was overlaid on the cell culture according to the product instructions for the cell support, and slowly turned with tweezers from the end of the support to recover the cell culture. The operation was not efficient because the holding condition after stacking was standing at 20 to 25 ° C. for 5 to 6 minutes. In addition, when turning the end of the support slowly with tweezers during the recovery operation, there are cases where the cell culture slips (flows away) from the support, and as a result, this may lead to damage. Some skill was required for operation. Furthermore, it was very difficult to peel off the cell culture during the operation of transporting and bringing it into close contact with the transplanted site and turning the end of the support with tweezers after standing.

  As described above, according to the device A of the present invention, a series of operations such as adsorption (collection, recovery), retention, transfer to the affected area, and application (transplantation) to the affected area of the cell culture (not shown). Can be performed very simply, quickly and reliably.

  The instrument A of the present invention includes a housing 1, a handle 2, a drainage operation unit 3, and the like, and in particular, a liquid absorption provided inside the housing 1 through the opening 13 of the holding unit 12 of the housing 1. The material 11 has a very simple structure utilizing the liquid absorbing action of the conductive material 11 and the surface tension of the absorbed immersion liquid 5, and the holding surface 121 of the housing 1 is made of a hydrophobic material, The transfer of the immersion liquid 5 to the affected part 7 is promoted, the cell culture 4 is very easily attached, and the operability is also good.

  As mentioned above, although the instrument of the present invention has been described based on the illustrated embodiment, the present invention is not limited to this, and the configuration of each part may be replaced with an arbitrary configuration having the same function. Can do. Also, any other component may be attached to the present invention.

  Note that the functions related to the drainage operation described based on the embodiments shown in the present invention are for supporting the convenience of the present invention, and are omitted or excluded when further convenience of the instrument is required. It is possible.

A ... Device 1 of the present invention ... Housing 11 ... Liquid absorbing material 12 ... Holding part 121 ... Holding surface 13 ... Opening 131 ... Opening wall (thickness)
14 ... drainage port 15 ... upper surface 16 ... groove 17 ... front surface 18 ... opening / closing part 19 ... normal 2 ... handle 21 ... shaft 3 ... drainage Operation part 31 ... movable plate θ ... inclination angle 4 ... cell culture 5 ... immersion liquid 6 ... container 7 ... affected part

Claims (13)

A device for collecting and delivering a cell culture released in a liquid, the cell culture holding part having a hydrophobic cell culture holding surface with at least one opening, and through the opening A liquid-absorbing part disposed on a side opposite to the cell culture holding surface of the cell culture holding part, wherein the opening is capable of sucking the liquid, and the opening is used to connect the cell culture to the cell culture. The cell culture is configured to have at least one contact with the cell culture holding surface when brought into contact with the holding portion, the cell culture holding surface is disposed on the outer surface of the instrument, and the cell culture is The instrument is held on the cell culture holding surface by the action of the surface tension of the liquid sucked into the liquid sucking part .
  The instrument according to claim 1, further comprising a hollow housing, wherein the liquid absorbing part is disposed inside the housing, and at least one outer surface of the housing is configured as a cell culture holding part.   The device of claim 1 or 2, wherein the cell culture holding surface is more hydrophobic than the site for delivering the cell culture.   The device according to any one of claims 1 to 3, wherein the cell culture holding surface has a porosity of 5 to 90% with respect to the area.   The instrument according to any one of claims 1 to 4, wherein the cell culture holder has a form of a single porous body, a porous body, or a mesh. The instrument according to any one of claims 1 to 5, wherein a distance between the cell culture holding surface at the opening and the surface of the liquid absorption part is 0.1 to 2.0 mm.   The instrument in any one of Claims 1-6 in which a liquid absorption part contains a liquid absorptive material.   The instrument according to claim 7, wherein the liquid-absorbing material includes one or more materials selected from the group consisting of a porous body, a nonwoven fabric, a polymer material, a pulp material, and a paper material.   The instrument according to claim 1, further comprising an operation handle. Further comprising a hollow housing, the housing, for performing a drainage operation of hydrated liquid-absorbent material, for distribution within the operable movable plate from the outside, according to any one of claims 7-9 Instruments. The instrument according to any one of claims 7 to 10 , further comprising a hollow housing, the housing having an opening / closing part for exchanging the liquid-absorbing material. A method for collecting and delivering a cell culture comprising:
(I) providing the device according to any one of claims 1 to 11;
(II) contacting the cell culture holding part of the device with the cell culture released in the liquid;
(III) a step of recovering the cell culture by adsorbing it to the cell culture holding unit together with the suction of the liquid by the liquid absorption unit;
(IV) moving the cell culture to a target site that is hydrophilic from the cell culture holding surface while adsorbing the cell culture to the cell culture holding unit;
Including said method.   The method of claim 12, wherein the target site is substantially free of moisture on its surface.

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