CN205035384U - Stem cell ball cutting collection device - Google Patents
Stem cell ball cutting collection device Download PDFInfo
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- CN205035384U CN205035384U CN201520553888.1U CN201520553888U CN205035384U CN 205035384 U CN205035384 U CN 205035384U CN 201520553888 U CN201520553888 U CN 201520553888U CN 205035384 U CN205035384 U CN 205035384U
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Abstract
The utility model discloses an apoptosis, sudden change, cell damage, nerve cord cell fifferentiation and operating time length and the uneven scheduling problem of neural ball size easily take place for the nerve cord cell that the solution nerve cord cell process of going down to posterity results in, provide a stem cell ball cutting collection device. A stem cell ball cutting collection device, the device includes stem cell ball cutterbar, stem cell ball receiving flask and have the malleation and the aspiration pump of the two -way function of bleeding of negative pressure, by the tube coupling between stem cell ball cutterbar and the stem cell ball receiving flask, this stem cell ball cutterbar has a body, screen cloth that two intervals of body internal fixation set up, and the distance between the two screens net is 18 -22mm. The utility model provides an in vitro culture nerve cord cell easily break up and exchange dead technological problem, reach the amplify purpose of people's nerve cord cell of long -term cultivation, the characteristic of people's nerve cord cell is stabilized moreover, the quality is controllable.
Description
Technical field
The utility model belongs to cell culture apparatus field, specifically, relates to a kind of stem cell ball cutting collection device.
Background technology
At culture of neural stem cells neural or neural precursor, tumor stem cell, during embryonic stem cell, usually occur dry cell mass or claim stem cell ball, the method separation all using trypsinase or trypsinase to add the isolated cell ball of EDTA of going down to posterity in long-term cultivation is gone down to posterity.Fractionation cell membrane acceptor due to these enzymes of life-time service produces damage in various degree, has a strong impact on the life-span of stem cell in long-term cultivation.The generation of nervous system disorders and development reduce due to nerve cell number or changing function causes.Related neurodegenerative disease can be treated by neural stem cells transplantation.Embryonic stem cell not only plays very important effect in the research fields such as mechanism of action, and also has extremely important effect in various diseases control.Cultivate tumor stem cell occur in research tumour and play an important role in control.So a kind of fast and convenient, the stem cell ball partition method of damaging cells is very unimportant.
Neural stem cell is a kind of multipotential stem cell, and under relevant cell factor stimulates, can breed in vitro and form special ball-like structure, this ball-like structure is called as neural ball.Neural ball has special microenvironment, can keep survival and the propagation of stem cell in ball, and gives the potential that neural stem cell breaks up to ectoderm cell (neurone and neurogliocyte).The preparation of neural ball is the committed step of neural stem cell treatment, and their quality directly has influence on the clinical treatment curative effect of neural stem cell.
Method conventional at present comprises enzyme digestion or mechanical piping and druming method.Enzyme digestion is that the method utilizing independent pancreatin or pancreatin and EDTA to share digests neural ball.Research display enzyme digestion can cause stem cell to be undergone mutation, and also there is the shortcomings such as operating time long and cell injury is large.Machinery piping and druming method adopts dropper or transfer pipet repeatedly to blow and beat tissue, although the operating time is short, same large to cell injury.And the cell mass size obtained is uneven, causes partial nerve sphere volume excessive, the survival of the stem cell that affects the nerves, growth and differ entiation.Have people to use sharp cutter patterning method recently, this method had large improvement than former method, but can cause a lot of cell debris, and the normal growth that these fragments may affect stem cell is grown.Therefore, set up a kind of easy, quick, without modificator gene sudden change, less and the preparation method of neural ball of uniform size can be prepared to cell injury, be current problem demanding prompt solution.
Utility model content
The utility model be solve that neural stem cell succeeding generations causes easily there is apoptosis, sudden change, cell injury in neural stem cell, neural stem cell differentiating and the operating time long and the uneven first-class problem of neural ball size, provide a kind of stem cell ball to cut collection device.
The utility model also provide having of utilizing said apparatus to carry out easy, quick, without modificator gene sudden change, cutting and separating propagating method to the less stem cell ball of cell injury.
The utility model solves the technical scheme that its technical problem adopts:
A kind of stem cell ball cutting collection device, this device comprises stem cell ball cutting unit, stem cell ball receiving flask and has the off-gas pump of malleation and negative pressure bidirectional air extracting function, connected by pipeline between stem cell ball cutting unit and stem cell ball receiving flask, this stem cell ball cutting unit has a body, body internal fixtion two spaced screen clothes, the distance between two screen clothes is 18-22mm.
As preferably, described screen cloth is stainless steel mesh, and the aperture of two screen clothes is respectively 350 ± 40 μm and 104 ± 20 μm, and the larger screen cloth in aperture is near stem cell ball receiving flask.Further, the aperture of two screen clothes is respectively 350 ± 10 μm and 104 ± 5 μm, and the size in aperture controls significant to the cutter of stem cell ball.
As preferably, this device is little stem cell ball receiving flask after also comprising air cushioning bottle, Y-tube, culturing bottle and cutting, off-gas pump is connected by pipeline with air cushioning bottle, air cushioning bottle is connected by pipeline with stem cell ball receiving flask, culturing bottle is connected with two interfaces of Y-tube by pipeline respectively with stem cell ball receiving flask, and the 3rd interface of Y-tube is connected with stem cell ball cutting unit.This structure is used for the large scale culturing of stem cell.
As preferably, the exit end of stem cell ball cutting unit is provided with the pipe connecting of a reducing.
A kind of stem cell ball cutting collection device, the stem cell ball cutting unit that this device comprises syringe and is connected with syringe, this stem cell ball cutting unit has a body, and body internal fixtion two spaced screen clothes, the distance between two screen clothes is 18-22mm.
As preferably, described screen cloth is stainless steel mesh, and the aperture of two screen clothes is respectively 350 ± 40 μm and 104 ± 20 μm, and the larger screen cloth in aperture is near stem cell ball receiving flask.
A kind of cutting and separating propagating method of stem cell ball, the method comprises the steps: that the stem cell ball cutting collection device described in a. employing carries out cutting and separating to stem cell ball: after the nutrient solution containing stem cell being sucked stem cell ball receiving flask, hydrodynamicpressure is utilized to make stem cell ball warp cell ball cutting unit passivity cut into little dry cell mass, then enter in culturing bottle, preset cell culture fluid in culturing bottle; B. the cell culture fluid containing little dry cell mass that a step obtains is carried out sub-bottle cultivation; When c, sub-bottle are cultivated, every 3-7 days changes a fresh cell medium, within every 20-30 days, repeats a step and carries out cutting and separating to stem cell ball.
As preferably, described stem cell ball is neural stem cell, each neural molecular biology is cut into 3-8 fritter with stem cell ball cutting collection device when reaching 500-2000 μm of size by the diameter of neural molecular biology, carries out 1:2,1:4 or 1:6 sub-bottle Secondary Culture.
Time ordinary method culture of neural stem cells neural (neural ball), with trypsinase or other collagenase, neural ball is digested to individual cells and carries out Secondary Culture.Once, along with passage number increases, the neural stem cell quantity of cultivation reduces (namely slowly dead) every 1 to 2 all Secondary Culture gradually.Conventional " tryptic digestion " method or be all difficult to neural stem cell (neural ball) Secondary Culture to surmount three months by " trypsinase+disodium ethylene diamine tetraacetate (EDTA) digestion " method or collagenase digestion.Analysis reason is as follows: 1. trypsinase can dissociate and destroy connection between each cell and slight damage cell-membrane receptor, can cut C and hold peptide section in cell, can the effects such as peptide is strong on the right side of protolysate arginine, lysine residue specifically.Use if a small amount of or once use the harm of trypsinase cell growth less, daughter cell even almost not had a significant effect, because cell self repair system works.If but repeatedly used trypsinization cell, would cause the infringement of cell to exceed the ability of cell self repair system, so the neural stem cell quantity of cultivating reduces gradually; 2. after neural ball being digested to individual cells with trypsinase, need again to stop tryptic activity with serum, and containing a lot of short cell differentiation factor in serum, so, traditionally during culture of neural stem cells neural, often run into a large amount of neural stem cell differentiating one-tenth neurone and spongiocyte (astroglia cell and oligodendrocyte), the neurone after differentiation is cultivated rear weather aging in vitro and is die, because neurone is thesocyte, it is acellular splitting function.Even if the astroglia cell after differentiation and oligodendrocyte can split into daughter cell and survive down, but under general condition, astroglia cell and oligodendrocyte reversely can not be divided into neural stem cell, more do not have the neuronic function of neural stem cell differentiating one-tenth.So according to a conventional method cannot long-term cultivation neural stem cell.
Can produce impact more or less to each cell by enzymic digestion isolated cell method, and " cell ball cutters " of the present utility model separate stem cells ball carries out Secondary Culture, to extreme portions cell without any infringement, but there is infringement or fatal to the small part cell that mechanical blades cuts to, the cell that this small part is fatal is die voluntarily, and change along with new and old nutrient solution, abolish along with old nutrient solution.This has no significant effect the new stem cell after going down to posterity.Therefore, cell ball cutters separate stem cells ball is the good method of culturing stem cells, is also one of the key factor of the utility model long-term cultivation expanding stem cells method in vitro.
The utility model solves vitro culture neural stem cell easily to be broken up and easily dead technical barrier, reach the object of long-term cultivation amplification human nerve stem cell, and the stability of characteristics of human nerve stem cell, quality controllable, for being widely used in clinically providing important, nexhaustible neural stem cell resource.The utility model people root is accordingly to neural stem cell cultured continuously time more than 3 years.In the utility model in the stem cell ball cutting collection device cutting and separating Secondary Culture experimental study that may be used for any cell ball and large-scale production.
Accompanying drawing explanation
Structural representation when Fig. 1 is the utility model stem cell ball cutting collection device suction stem cell ball;
Fig. 2 is the structural representation of the utility model stem cell ball cutting unit;
Fig. 3 is the structural representation of the utility model stem cell ball cutting collection device when cutting neural ball;
Fig. 4 is the another kind of structural representation of the utility model stem cell ball cutting collection device;
In figure: 1, off-gas pump, 2, air cushioning bottle, 3, stem cell ball receiving flask, 4, Y-tube, separate lines, 5, culturing bottle, 6, stem cell ball cutting unit, 7, screen cloth, 8, pipe connecting, 9, little stem cell ball receiving flask after cutting, 10, syringe.
Embodiment
Below by specific embodiment, and by reference to the accompanying drawings, the technical solution of the utility model is described in further detail.Should be appreciated that enforcement of the present utility model is not limited to the following examples, any pro forma accommodation make the utility model and/or change all will fall into the utility model protection domain.
Embodiment 1:
A kind of stem cell ball cutting collection device as depicted in figs. 1 and 2, this device is made up of little stem cell ball receiving flask 9 after stem cell ball cutting unit 6, air cushioning bottle 2, Y-tube 4, culturing bottle 5, stem cell ball receiving flask 3, cutting and the off-gas pump 1 with malleation and negative pressure bidirectional air extracting function.
Connected by pipeline between stem cell ball cutting unit and stem cell ball receiving flask, this stem cell ball cutting unit has a body, the spaced screen cloth 7 of body internal fixtion two.Screen cloth is stainless steel mesh, and the aperture of two screen clothes is respectively 350 μm and 104 μm, and the screen cloth of 350 μm is near stem cell ball receiving flask, and the distance between two screen clothes is 20mm.The exit end of stem cell ball cutting unit 6 is provided with the pipe connecting 8 of a reducing.
Off-gas pump is connected by pipeline with air cushioning bottle, air cushioning bottle is connected by pipeline with stem cell ball receiving flask, culturing bottle is connected with two interfaces of Y-tube by pipeline respectively with stem cell ball receiving flask 3, and the 3rd interface of Y-tube is connected with stem cell ball cutting unit.
The use procedure of the utility model device is as follows:
1, stem cell ball collection process: as shown in Figure 1, close the switch that Y-tube connects to stem cell ball cutting unit, transfer pipet inserts in the culturing bottle 5 containing stem cell ball, open off-gas pump, utilize negative pressure to make stem cell ball in culturing bottle with in liquid whole suction stem cell ball receiving flask 3, the many bottles of nutrient solutions containing neural molecular biology can be collected.
2, stem cell ball cutting collection process: as shown in Figure 3, stem cell ball in stem cell ball receiving flask is cut and collection process, open the switch that Y-tube connects to stem cell ball cutting unit, also close the path that culturing bottle 5 enters stem cell ball cutting unit simultaneously, open malleation off-gas pump switch, utilize malleation, make to circulate to stem cell ball cutting unit direction containing neural molecular biology in stem cell ball receiving flask 3, off-gas pump malleation is utilized to make stem cell ball with the stainless steel mesh of liquid quickly through passivity, start first by the screen cloth in 350 μm, aperture, again by the screen cloth in 104 μm, aperture, reach the object that large neural molecular biology becomes nervelet stem cell ball.This device is applicable to the use of large scale culturing neural stem cell.
Embodiment 2:
Stem cell ball cutting collection device as shown in Figure 4, the stem cell ball cutting unit 6 that this device comprises syringe 10 and is connected with syringe, this stem cell ball cutting unit has a body, the spaced screen cloth 7 of body internal fixtion two.Syringe 10 volume is 100-200ml, and the exit end of stem cell ball cutting unit 6 is provided with the pipe connecting 8 of a reducing.Described screen cloth is stainless steel mesh, and the aperture of two screen clothes is respectively 350 μm and 104 μm, and the distance after the close cutting of screen cloth that aperture is less between little stem cell ball receiving flask 9, two screen clothes is 18mm.This device is applicable to the use of laboratory or small-scale culture of neural stem cells neural.
First syringe is sucked the 100ml to be cut nutrient solution containing stem cell ball, utilize the thrust of artificial hand, make 100ml contain the nutrient solution of stem cell ball quickly through stem cell ball cutting unit and pipe connecting, enter the rear little stem cell ball receiving flask 9 of cutting.
Above-described embodiment is one of the present utility model preferably scheme, not does any pro forma restriction to the utility model, also has other variant and remodeling under the prerequisite not exceeding the technical scheme described in claim.
Claims (6)
1. a stem cell ball cutting collection device, it is characterized in that: this device comprises stem cell ball cutting unit (6), stem cell ball receiving flask (3) and has the off-gas pump (1) of malleation and negative pressure bidirectional air extracting function, connected by pipeline between stem cell ball cutting unit and stem cell ball receiving flask, this stem cell ball cutting unit has a body, body internal fixtion two spaced screen clothes (7), the distance between two screen clothes is 18-22mm.
2. stem cell ball according to claim 1 cutting collection device, it is characterized in that: described screen cloth is stainless steel mesh, the aperture of two screen clothes is respectively 350 ± 40 μm and 104 ± 20 μm, and the larger screen cloth in aperture is near stem cell ball receiving flask.
3. stem cell ball cutting collection device according to claim 1, it is characterized in that: this device also comprises the rear little stem cell ball receiving flask (9) of air cushioning bottle (2), Y-tube (4), culturing bottle (5) and cutting, off-gas pump is connected by pipeline with air cushioning bottle, air cushioning bottle is connected by pipeline with stem cell ball receiving flask, culturing bottle is connected with two interfaces of Y-tube by pipeline respectively with stem cell ball receiving flask (3), and the 3rd interface of Y-tube is connected with stem cell ball cutting unit.
4. stem cell ball cutting collection device according to claim 1, is characterized in that: the exit end of stem cell ball cutting unit (6) is provided with the pipe connecting (8) of a reducing.
5. a stem cell ball cutting collection device, it is characterized in that: the stem cell ball cutting unit (6) that this device comprises syringe (10) and is connected with syringe, this stem cell ball cutting unit has a body, body internal fixtion two spaced screen clothes (7), the distance between two screen clothes is 18-22mm.
6. stem cell ball according to claim 5 cutting collection device, it is characterized in that: described screen cloth is stainless steel mesh, the aperture of two screen clothes is respectively 350 ± 40 μm and 104 ± 20 μm, and the larger screen cloth in aperture is near stem cell ball receiving flask.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201520553888.1U CN205035384U (en) | 2015-07-28 | 2015-07-28 | Stem cell ball cutting collection device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201520553888.1U CN205035384U (en) | 2015-07-28 | 2015-07-28 | Stem cell ball cutting collection device |
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| CN205035384U true CN205035384U (en) | 2016-02-17 |
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| CN201520553888.1U Active CN205035384U (en) | 2015-07-28 | 2015-07-28 | Stem cell ball cutting collection device |
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- 2015-07-28 CN CN201520553888.1U patent/CN205035384U/en active Active
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