JPH05509292A - tissue implantation method - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Implantation □ Technique 1 No and No The present invention generally relates to the implantation and re-implantation of mammalian tissue in mammals. on how to More specifically, the present invention aims to improve the effectiveness of implantation. Sexual enhancement procedures; treatment methods including implantations; The present invention relates to tissue compositions useful for tion.

m people two plates The chorioallantoic membrane (CAM) is a highly vascularized extraembryonic fetal membrane in birds. it's a bird Provides gas exchange essential for embryonic development. It is difficult to obtain and proximally separate from the embryo. Because of its ease of use, it has been studied and utilized from both developmental and tumor biology perspectives. It is.

At the beginning of this century, CAM was used to study the growth of small tumor-transplant tissue. It was commonly used (Xarnofsky et al., New York Acad Sci ( 1952) 55:313). More recently, it has been used in membrane transport, gas exchange, and It has been used to study angiogenic activity and angiogenic activity.

In developmental biology, CAM is a protein that is associated with various types of avian liver tissue and organ primordia. It is also used as a substrate for growth (Ravles, Ann NY A cad Sci (1952) 55:302) o Is it an IFF chicken? Ebert cultured organ explants (Exbran (g) to study the effect of explants on corresponding tissues of developing eggs. (ProcNat Acad (1954) 40:337; J Nat Cancer In5t (1960) S. 73). Elur st et al. showed that some normal human tissues grown on CAM exhibited mitotic activity. reported (Austr　J　Exo　Biol　(1939)　17+　215 ). However, adult endothelial tissue survives for more than 3 days when implanted in CAM. There are conflicting reports that CAu5prunk & Fol cannot survive. kman, Virchows Arch B Ce1l Path (], 97 6) 21:31).

Implantation as a treatment method has found many applications in the medical field. ing. However, the widespread use of implantation is always limited by the survival effectiveness of the tissue.

For example, hair transplantain is used to treat alopecia (hair loss). There is. Baldness can be treated by surgically transplanting active hair follicles to the scalp.

The best reported report suggests that Parkinson's syndrome is caused by the autologous transfer of adrenal medullary tissue to the striatum of the brain. Although it has been shown that it can be improved by in-branch conversion, Suvlanted tissues survive for a relatively short period of time (Ding, Y., J. , et al., ChinMed J, (1988) 101:631: Hansen, J, T. et al., ExNeurOl (1988) 102:65; Strombe rg, 1. et al., Ex Brain Res (1985) 60:335).

(Recently, Brundln et al. (EX Brain Res (1988) 70:192), human fetal dopamine neurons were used in the same manner as in rats. Lindvall et al. (Arch Neu reported) .

Implantation is an important step in the early development of mammalian embryos. Imbi Toro fertilization techniques are available for immediate implantation and cryogenically delayed implantation. (Camus, M, et al. and 5teril (1989>51:460) a However, once again it has been proven that there are limits to the effectiveness of implantation. (Liu.

H, C, et al. Fert and Teril (1988) 49:649) o’1lieraer et al. (murt and 5teril (1989) 52:503) cultured human zygotes together on a monolayer of fetal bovine uterine fibroblasts. It has been reported that the effectiveness of implantation increases if the patient is nourished.

Finally, gene therapy may be used to treat various metabolic deficiencies and improve long-term improvement of proteins. It has been proposed as a treatment for diseases requiring administration (Green, H, et al. , Proc Nat Acad Set (1979) 76:5665; lmer et al., Proc Nat Acad Sci (1987) 84:10 55).

But once again, successful re-implantation of genetically treated tissue showed that the technology has limitations.

1 ministry, 1 statement One embodiment of the invention provides treatment for a mammal in need of tissue implantation. A method of treatment, which includes: That is, development with chorioallantoic membrane (CAM) A live amniote egg is prepared; it does not produce a domesticated mammalian tissue consisting of a population of cells. implanting mammalian cells or tissues in or on the CAM to Remove the conditioned tissue from the CAM by excisional means; This involves implanting the tissue into mammals.

In other embodiments, developing chicken eggs of the genus Wildfowl are used; In an embodiment, developing ostrich eggs of the genus Ostrich are used.

In other embodiments, the invention provides treatments for treating genetic and metabolic deficiencies in gene expression. used for medical treatment. In yet other embodiments, the invention provides treatments for treating alopecia. used for medical treatment. In other embodiments, the invention provides a method for treating Parkinson's disease. used for treatment.

In yet other embodiments, the present invention provides for mammalian plain plantations. Used to facilitate implantation of the embryo into the recipient mother's uterine epithelium be done. In another embodiment, the invention provides methods for fertilizing mammalian oocytes in vitro. provide a method to do so. In a final embodiment, the invention relates to mammalian implants. composition of tissue useful for avian CAM; Consists of mammalian tissue and growth factors.

1 plate of main soup stock Kusenkun Steady state Implants of the ``r same mx system'' are obtained from living organisms other than the recipe end. contain cells or tissues with different genetic compositions other than the host tissue. be caught. “Autologous” grafts include cells or cells that originate from the recipient organism. Tissues (with the same genetic composition) are included. Finally, in "homotypic" grafts contain cells or tissues of the same genetic composition and of the same type and function. be caught.

“Enlarged” tissue is defined as tissue that has substantially increased in size as a result of mitotic proliferation. That is.

"Amniotic eggs" have a shell and provide their own water supply when expelled from the parent's body. retains salary. It contains egg yolk and rich albumin. the embryo is inside the egg They grow up outside of their parents. Examples include birds and reptiles.

“Acclimated tissue” is grown on the CAM tissue and as a result enlarges and becomes substantially bloodless. Vascularization takes place.

A "developing" egg is one that begins ontogeny in response to appropriate incubation conditions. This is a fertilized amniotic egg.

“Fertilization unit” refers to a mammalian embryo that is normally at the stage of blastocyst development; It consists of mammalian uterine epithelial tissue that is runt. In-branching of the embryo - prior to dialysis The uterine epithelial tissue is found in developing amniotic eggs, such as chicken or ostrich eggs. Implanted under the choroidal epithelium of the CAM. The fertilization unit is essentially re-implanted into the uterine epithelium of the mammalian mother.

Implantation involves moving tissue from one area to another. It is to bury the mobile tissue so that it continues to live. Tissues are derived from the organism from which they are derived. Can be imbued with new locations or disparate recipe ends. Imbunted Organizations can be located within organizations of the same function or different functions.

A subject who is “in need” of medical or surgical treatment will be improved by that treatment. An individual with a trauma, disease, defect, or medical condition that may cause physiological processes (e.g. implantation of an embryo during gestation) It is an organism that is promoted by treatment and function.

“Neurological defect” means an external defect in nerve cells or tissue that results in loss of nerve function. It is a wound, disease, or degeneration.

The "structural sequence" of DNA hoods the polypeptide.

“Transplantation” means that the organization to be moved is the same organization in another location. This is one form of imbranching that is combined with a mold. Trans Plantae If an action is a movement from one part of an individual to another, it is a homotamination. It is ``self-transplantation'' which is equivalent to Ipuinbranch-Sikong.

"Vascularized" tissue refers to tissue that develops new blood vessels that connect with the host's circulatory system and become patent. .

day ≦, Some traumas, diseases, and defects are caused by tissue implantation or trauma. Can be treated by lance plantation. In addition, mammalian pregnancy-like Several physiological processes involve this tissue implantation. these The effectiveness of treatment and the success of these methods depend on the persistence of the implanted tissue. The usefulness and direct related to Disclosed herein is the effectiveness of implantation. This is a method of acclimatizing the tissue in order to increase the treatment of mammals in need of treatment and the use of effective regimens for such treatment. This is the composition of the weave.

Briefly, the method of tissue acclimation and subsequent use in treatment is based on the chorioallantoic membrane. preparing an amniotic egg having an embryo that has developed (CAM); and mammalian cells or Implanting tissue onto or into the CAM. As the embryo continues to grow Therefore, the tissue grows on or in the CAM, which is connected to the egg's circulatory system and the implant. patency has been established between the vasculature of the embryo and the embryonic environment is nourishing and growing. This is because it is rich in factors. Additionally, the graft tissue responds to embryonic angiogenic factors. It becomes rich in blood vessels. The conditioned mammalian tissue is then placed along the sticky portion of the CAM. Extracted and injected into the mammalian host in the optimal location for that particular treatment method. will be played. Compared to direct braided implants, this implant The effectiveness of this treatment is determined by the increased vascularity of the implanted tissue and the formation of sticky embryonic vessels. The tendency to rapidly form functional connections with host tissue capillaries greatly improves It was. Furthermore, growth factors provided by the liver tissue are also present in the graft and in the donor tissue. can facilitate integration into lance plantation sites.

Although this method may utilize any amniotic egg in a preferred embodiment, ・Insect nets or seaweed eggs are used. In a more preferred embodiment, wild chickens One member of the genus Chickensis is used. It has been tested and proven to be pathogen-free. Fertilized chicken eggs can be easily obtained commercially. artificial incubation tion methods are known to those skilled in the art. In other preferred embodiments, Beta power of the genus Ostrich is used. Eggs that are proven to be free of pathogens are Ostrich and its eggs are not available commercially, but ostriches and their eggs are commercially available. g Sky 0strich Ranch, Plentywood, Mon (tana), which meets the same standards currently used for chicken eggs. power) and power) can be monitored. These methods can be used by those skilled in the art (in general). This is publicly known. Artificial incubation methods for ostrich eggs are known ( Jarvis9M1. F., 0strich (1985) 56:42).

Techniques for preparing CAMs that accept chicken eggs and tissue grafts (well known to those skilled in the art) (For example, Zwilling, E, Trans 1ant B Sho, ( 1959) 6+115). The same technical ability (for different gestational periods) It may also be applicable to amniotic eggs of other species with appropriate changes. Mammalian phosphatide (A) prolongs graft survival and prevents subsequent re-incubation. Modified to allow for lunch-time conversion. This modified technology can be used to Incubate in a humidified chamber (incubator) for 6 to 9 days prior to drying. Incubated and maintained at °C.

The age of the chicken embryo determines the amount of mammalian tissue that should be implanted. transplant The more tissue there is, the more blood vessels the CAM surface and the chicken heart pump. Requiring increased capacity, this also provides better vascularity.

Locate the space at one end of the egg by pointing a small, low-intensity light source through the egg. ) and make a small (1 mm) hole so that air can enter this space. Let it come in. Next, identify the embryonic and non-embryonic membranes and draw a pencil in the center of the space. Mark the position by writing “°X”. Use a small triangular file to mark the line. This creates a 1cm square window around the "X" on the shell. like this The groove created in the uterus gradually becomes deeper and deeper beneath the uterine membrane. The surface of this egg is 80 Wipe the area around “X” clean for a few seconds with a cotton swab containing % ethanol solution, and then Let the surface dry. Next, insert the window into one corner of the square using sterile watchmaker's forceps. Insert the tip of the egg, lift it up to expose the egg membrane underneath, and remove.

Empty the egg membrane by carefully pushing the tip of the sterile watchmaker's forceps. Let the qi enter and allow the CAM to separate and ``fall''. This membrane is then coated with a small Make an incision with a scalpel and carefully peel it away from around the window to expose the CAM. remove.

Observe the surface of the CAM at low magnification using a dissecting microscope to identify major blood vessels. child blood vessels trace their origin to points of convergence that form the allantoic vascular supply to the embryo. It is something. Vascular harvesting on CAM surfaces for mammalian tissue implantation A tentative site is established near the calculus, and the choroidal epithelium is injected with a sharp glass needle tip. Gently peel away one or more graft sites.

Temporarily block this window by applying several strips of 5cotch wide tape to the opening. ingredient.

Ensure that live tissue grafts from deeply anesthetized mammals are maintained under sterile conditions. Ru. A small piece of living tissue (not exceeding 1 mm x 3 mm xll) is quickly removed. Take it out and secure it. This tissue was confirmed in advance as a CAM in-branch-shicon site. Place the sterile culture solution in a chilled (4°C) vitri dish immediately. Immediately transfer to liquid.

This chilled solution lowers metabolic demand and reduces the amount of damage induced by trauma on the CAM. causing the diffusion of leukotrienes and proteins that cause a local inflammatory response. This is favorable for the survival of cells in tissues.

The tissue is then made a final incision, the surrounding connective tissue is removed, and the It is cut down to the desired size. This step allows the microvasculature to be exposed on the surface of the graft. This is important for rapid fusion of components and choroidal vessels. Regarding this, This will be discussed further below.

Next, remove the tape, open the egg, and insert the imp that you checked earlier. Place the plantation site below the 6th part of the window fabric while slowly rotating the egg. Determine the position. Place the small piece of tissue to be implanted in a vetri dish and gently use sterile forceps ( remove it by rubbing it under each piece and pulling it away from the culture medium.

Excess culture medium can be quickly wiped off by placing the forceps on sterile blotting paper. and then transferred to the CAM leaving a minimum amount of residue on its surface. .

Gently place the tissue on the surface of the CAM implantation site and without introducing additional bleeding from the small capillaries running beneath the choroidal surface. , carefully moving it to the previously stripped surface, ensuring good contact.

In situations where excessive amounts of culture medium remain on the CAM graft surface, use a sterile cell Use the tip of a loin sponge (e.g. Ti! Shiko Paper) to squeeze the liquid inside. Eliminated it by removing it.

For rapid fusion of choroidal vessels with microvascular components exposed on the graft surface. Various methods may be utilized to improve the "acceptance" of the CAM, as appropriate. . In addition to surface tearing of the CAM, to reduce the time required for vessel fusion, (In 　addftfanal　to　5upperificial　1aceratio n of the CAN 5urface fr+.

rder to decrease the titse required for vascuLtr fusion,).

Critical to graft survival is the rapid fusion of blood vessels, which allows Blood is provided by the circulatory system of the embryo inside.

This fusion is achieved within minutes by the microvascular components in the fabric, which Check for exposed microvascular components of the explant. The forces involved in this cognitive process Although little is known about the choroidal epithelium, it is It depends in part on the superficial position of the coelomic blood vessels running through it.

Furthermore, the location where the detached choroidal epithelial cells come into contact with the implanted tissue It is possible that the vascular components of the underlying choroid may be moved away from the choroid. It will be exposed.

Vascular fusion is sometimes performed using a small (5 mm x 5 mm) sterile clear plastic tube. Transplanted using “Chip J” (Gel-Bond, FMCCarp,) manufactured by It is also preferably carried out by covering a piece, whose weight and hydrophilic surface This is because it prevents the graft from drying out and at the same time promotes contact with the CAM.

After implantation, the egg window is sealed with tape and its edges are heated with a hot scalpel. Seal by applying paraffin wax using the flat part of the this The eggs are then incubated at 37°C. The graft is placed under a stereoscopic microscope. Place the egg down, remove the window tape, and rotate the egg so that the graft is visible. Therefore, it can be observed. By using a stereomicroscope, this and previous steps It is also possible to take photos. If high magnification is used, 1 post-implantation Check blood flow in the superficial small vessels that develop at the graft-CAM interface within 2 days. By this, it is possible to determine the viability of this graft. like that Blood vessels continue to increase in size over time, and some types of tissue (i.e., adrenal medulla) can be distinguished by their high degree of vascularization.

Re-implantation of the conditioned graft into a mammalian host anesthetizes the recipe end and remove the intact microvessels using surgical techniques known to those skilled in the art. It is done by exposing the part of the target organ that has. In the case of cerebrum, please contact a person skilled in the art. Stereotactic neurosurgery allows tissue to be placed in the appropriate part of the brain with minimal damage. Delivered.

The grafts should be cooled (still in the egg) by first cooling for a few minutes in the refrigerator. ), ready for re-implantation. Subsequently, the shell window is opened, and approximately 1~ Two cubic centimeters of chilled culture medium is placed inside to moisten the graft surface. . After 20-30 seconds, the culture medium is aspirated to expose the graft and the underlying choroid. Peel off carefully.

In a preferred embodiment, the graft is then carefully inserted with a large gauge hypodermic needle. By applying deep, controlled “suction”, the remaining allantoic membrane can be removed from the opening surface. be taken out. The hypodermic needle has a modified tip with a sharp, rounded, non-beveled opening. It was made by Such suction is internalized to create a vacuum. A close-fitting “plunger” that joins a second needle Executed by

The aspirated tissue remaining in the lumen of the larger needle shaft (mantle) is then The implant post is safely transferred to the re-implantation site and the indicated tissue is placed on or in a surface. An alternative embodiment is to demarcate the graft. It is removed by making a large incision, and then cutting out the surrounding tissue.

Needless to say, all of the above re-implantation procedures are performed using sterile techniques. It is essential to carry out the following. However, a special resistance to infection is engineered into the egg. It should also be mentioned that there may be Ru. This has not been reported by other researchers so far. subordinate Therefore, it is possible to achieve graft survival in ovo without maintaining complete sterility. It is Noh. On the other hand, similar resistance to infectious agents can be reintroduced into mammalian hosts. This is less likely to occur in tissues that have been exposed. Therefore, re-inbranch It is essential to take maximum precautions when dealing with

The examples given below are intended to be illustrative only and are not intended to limit the invention. isn't it.

Actual "E example" - in Alopecia (hair loss) is currently diagnosed using skin tests from areas of the skin where hair grows in patients with alopecia. Treatment is done by removing the hair and implanting them into the hairless area. There is. This method is useful for patients with alopecia who have a constant atrophy of the skin where hair grows properly. Limited due to Zabar.

Dermal papillae contain cells that can induce hair growth, and are produced from the cells of the dermal papillae. An unknown growth factor must be continually supplied to maintain hair growth. baldness The head contains papillae of specialized hair follicles that give rise to the cells needed for hair growth and maintenance. It is caused by hair loss due to prolonged inactivity of certain stem cells. herein The disclosed method involves the use of hair follicles derived from the dermal papilla of active hair follicles on or in the CAM of chicken eggs. It includes a step of synthesizing stem cells. By proliferation of these stem cells on CAM , a reservoir of highly activated autologous stem cells may be generated that can be transplanted into bald areas. provides a large amount of tissue.

This autologous transplantation tissue is implanted under the inactive hair follicle and subsequently on the endogenous stem cells of the dermal papillae and on the associated keratinocytes. It exerts a stimulating effect, resulting in hair growth.

History of branch blood In humans 1° Techniques for in vitro fertilization generally include a donor (which is also a recipe) Collect oocytes from the end (which can also be endo) and inject at least one oocyte. Fertilize in vitro, and then recipe it on or into the mother's uterine epithelium Including the step of re-implanting. One of the most difficult aspects of this procedure is the reintroduction. This is a plantation process. Only 1 out of 6 (or more) trials It becomes a fetus and waits for the day of delivery.

To improve the effectiveness of re-implantation, the fertilized eggs are first placed on the CAM. inject into the growing uterine epithelial graft tissue (harvested from the mother at the end of the recipe). to After implantation and establishment of the embryo in this epithelial graft, all fertilized units are Re-implant into mother body.

Briefly, small explants of uterine epithelium are removed from the uterus prior to or at the time of egg retrieval. ``Stroma-down'' is obtained on the CAM. J multiply. During proliferation on the CAM, the uterine epithelium, to receive the fertilized blastocyst. Can be prepared by hormonal regulation. After in vitro fertilization, mammalian embryos are extracted from the uterine tissue. Transfer to the surface of the explant and implant. Implantation is to be inspected This can be confirmed further. Confirm proliferation and transplantation of the entire fertilized unit to its highly vascular Microsurgically re-implant the mother's uterine wall, including the newly generated stroma. and return it.

Because it is possible to adjust the epithelium without performing systemic hormone therapy on the recipe end. therefore, the effectiveness of liver transplantation is increased. Furthermore, inserting the embryo into the recipe end for successful implantation This has the advantage of being able to confirm the situation before it occurs, thereby avoiding unnecessary patient trauma. Wear.

Shigo Shiyu In Human X In another embodiment, the uterine epithelial tissue is first cultured on the CAM and then the blastocyst Instead of implanting the zygote (fertilized oocyte) into this cultured tissue, The CAMs are cultured in small plastic enclosures within the embryonic body cavity. Advantages of this method is that the total fertilization unit can be kept small (<3 mm), which means that the final improving the efficacy of in vivo re-implantation.

Fruit "1 row" d In humans In another embodiment, the oocytes are transferred directly to the CAM rather than in a culture dish or test tube. fertilize on top. A 2mm hole covered with a 3mm gold screen with a diameter of approx. 7) sterilized transparent hydrophilic plastic chip (Gel Bond, F MCCarp, ). The choroidal epithelium was removed using a newly polished glass rod. Remove it from a 21 nm x 2 mm area by gradually peeling it off without causing bleeding. with money Place the plastic chip with the covered hole on the peeled part on the CAM and close the hole on the body. Fill with cavity fluid (no blood).

The oocyte is then placed in the hole along with the cumulus (coronal) cells attached to it. oocyte Treatment with hyaluronidase dramatically reduces the number of coronary cells during cyst maturation to expose the oocyte surface. After the oocytes have matured, sperm is added and received. Incubate with oocytes for fertilization.

This method requires that the embryonic environment of the CAM is more effective than the in vitro environment for successful fertilization and fertilization. It has the advantage of being more effective for the survival of goko. This is true in vitro. increasing the overall effectiveness of the fertilization procedure.

Support ball In this embodiment, a large number of To acclimatize the autologous adrenal medulla-derived tissue used for brain transplants, As described above, a method is used in which mammalian tissues are cultured on avian CAM. even here The main reason limiting the success of this procedure is the lack of survival of the graft at the insertion site. There is a problem that (see example Ding, Y, J, et al., Chin Mad J ( 1988) 101:631; Hansen, J. T., et al.

(1988) 102:85). Expranchysilane and striae of the adrenal medulla Surgical techniques for in-branch surgery of the body are known to those skilled in the art (same as described above).

Transplantation using standard protocols for immediate transplantation of adrenal tissue. An important process involving the fusion of capillaries at the surface of plant tissue with capillaries in the cerebrum. The initial steps are never performed fast enough and on a large enough scale to support tissue survival and proliferation. It won't happen. Increasing the number of surface capillaries during CAM implantation and also by acclimating and proliferating the parenchymal tissue itself. Transplant survival after re-implantation into the site was significantly increased. It will be done.

Fruit] 111 in Parkinson's In this embodiment, the implant is placed in the striatum of a patient suffering from Parkinson's syndrome. The tissue to be used is derived from human fetal tissue. More specifically, the cerebral tissue of a miscarried baby Transplanting dopamine-producing neurons from the ventral midbrain region onto the CAM of avian embryos. do. After 5 to 10 days, the conditioned fetal tissue was removed from the CAM and Implant into the striatum of patients suffering from the syndrome.

Stereotaxic brain surgery ensures that the surgery is performed in the appropriate position. These techniques are well known to those skilled in the art. It is. Patients with cyclosporine, azathioprine, and steroid therapy treatment to minimize the risk of transplant rejection.

1 craftsman knee on the car at ° Gene transfer of naturally occurring gene products using gene transfer techniques known to those skilled in the art. Diseases characterized by deletions (e.g. diabetes and insulin) or of gene products Diseases characterized by an improved response to induction (heart disease and tissue plasminogen activation) The gene is inserted into the autologous body cells of a patient suffering from the disease.

The choice of the type of somatic cell into which the therapeutic gene is inserted depends on the desired mode of delivery. For example, transduced epithelial keratinocytes are determined to have a gene product transduced endothelial cells are used to provide a “dermatitis” in which introduced into a vascular graft to provide secretion of the gene product directly to the circulatory system; and Transduced parenchymal cells (such as hepatocytes) provide organ-specific delivery of gene products .

In each case, genetic treatments were performed (e.g., transformation or transduction). cells) are first introduced into the CAM of a developing avian embryo; to loEI, or for 10 to 20 days in the case of ostrich eggs. This culture During the incubation period, the number of introduced cells increases, forming tissues and then vascularizing . This adapted tissue is removed and placed in the desired location within the patient's body using the techniques described above. Re-implant with homotype.

Genetically modified autologous cells may also be used in combination with other methods of treatment mentioned above. It can be done. For example, adrenal medullary cells are isolated prior to implantation onto the CAM and in the park. Before reinplanting the cerebrums of patients with cancer Can be genetically modified.

Essential m The present invention provides a method for treating mammals in need of tissue implantation with chorioallantoic membrane (CAM). ) Prepare developing amniotic eggs and inject mammalian cells or tissues onto the CAM. The conditioned mammalian tissue is produced by blunting and the conditioned tissue is removed from the CAM by excision means. by removing the tissue and re-implanting the conditioned tissue into the mammal. A method of treating is disclosed. The present invention also discloses related compositions and methods of treatment. do.

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Claims (28)

[Claims] 1. Persons preparing mammalian tissue for implantation into mammalian subjects A method comprising: providing a developing amniotic egg having a chorioallantoic membrane (CAM); Mammals on or in the CAM to produce conditioned mammalian tissue containing cell populations implanting a cell-like cell or tissue, and removing the conditioned tissue from the CAM by extraction means. . 2. Claim 1, wherein the mammal in need of implantation is a human. Method described. 3. 2. The method of claim 1, wherein the amniotic eggs are from bird nets. 4. 4. The method of claim 3, wherein the eggs are from the genus Wildfowl. 5. 4. The method of claim 3, wherein the eggs are from the genus Ostrich. 6. 2. The method of claim 1, wherein the extraction means is suction. 7. 2. The method of claim 1, wherein the extraction means is an incision. 8. 2. The method of claim 1, wherein the tissue is autologous. 9. 9. The method of claim 8, wherein the tissue is homotypic. 10. 8. The genome of a subpopulation of cells in said tissue is genetically altered. The method described in. 11. 9. The genome of a subpopulation of cells of said tissue is genetically altered. The method described in. 12. The tissue implantation results in polypeptide synthesis in a subpopulation of cells. and wherein said genetic alteration is a construct encoding said polypeptide. DNA containing a structural sequence is inserted, and the structural sequence functions as a control sequence of the DNA. 12. The method according to claim 11, comprising being coupled so as to. 13. The claim that the synthesis of said polypeptide is necessary for alleviating a genetic defect. The method according to item 12. 14. said tissue implant is for the purpose of treating alopecia; The cells implanted in the CAM are derived from the dermal papilla of an active hair follicle of the subject. the emulsified tissue is comprised of native stem cells, and the emulsified tissue is desired for hair growth in the subject. 9. The method of claim 8, wherein the method is implanted into the skin at a site where the implant is performed. 15. The tissue implantation is intended to treat Parkinson's disease. and the conditioned tissue is implanted into the striatum of the subject's brain. , the method according to claim 8. 16. 16. The method of claim 15, wherein the tissue is derived from the subject's adrenal medulla. 17. 2. The method of claim 1, wherein the implanted tissue is allogeneic. 18. The tissue implant is intended to treat Parkinson's disease and said tissue is derived from human fetal tissue and is implanted into said subject's cerebral striatum. 18. The method of claim 17, wherein the method is runt. 19. The tissue implant is intended to treat neurological damage 9. The method according to claim 8. 20. said tissue is derived from neural tissue of said subject and is implanted at said injury site. 20. The method of claim 19, wherein the method is runt. 21. A method of implanting a mammalian embryo into the epithelial tissue of the uterine wall of a mammalian mother, the method comprising: , the method is Developing amniotic eggs with chorioallantoic membrane (CAM) are prepared and uterine epithelial tissue from the mother is prepared. The epithelium is conditioned by implanting an explant on or into the CAM for a predetermined period of time. and placing the mammalian embryo on or in the conditioned epithelium; The embryo and the conditioned tissue are incubated to form a fertilization unit until engraftment in the tissue. remove the fertilized unit from the CAM and inject the unit into the mother's in vivo uterine epithelium. implanting and restoring the position, A method that encompasses. 22. 22. Used in combination with in vitro fertilization of said eggs. Method described. 23. Before the fertilization unit is implanted into the uterine epithelium of the mother, 1 22. The method of claim 21, wherein the method is incubated for a period of more than 20 days. 24. A method of implanting a mammalian embryo into the epithelial tissue of the uterine wall of a mammalian mother, the method comprising: , the method is Developing amniotic eggs with a chorioallantoic membrane (CAM) are prepared and mammalian blastocysts are placed in the CAM. the blastocysts are incubated for a predetermined period of time; The nuclear blastocyst is removed from the AM and the blastocyst is transferred to the mother's in vivo uterine epithelial tissue. to be implanted in A method that encompasses. 25. 1 day before the blastocyst is implanted into the uterine epithelium of the mother. Incubated in or on said CAM for a period of more than 20 days. The method according to item 24. 26. A method of fertilizing a mammalian oocyte, the method comprising: Developing amniotic eggs with a chorioallantoic membrane (CAM) are prepared and the chorioallantoic epithelium of the CAM is Transparent hydrophilic plastic with holes partially peeled and covered with gold screen Prepare chips made of The chip is placed on top of the stripped epithelium and at least one mammalian oocyte is placed on top of the peeled epithelium. place in a hole, introduce sperm of the same species into the hole, and incubating the oocytes and sperm together for a predetermined period of time; A method that encompasses. 27. Claim wherein the oocyte is treated with hyaluronidase prior to introduction of sperm. 26. The method described in 26. 28. A composition of tissue useful for mammalian implantation, the composition comprising: but with mammalian tissue intended to be implanted into a mammalian host, and with avian CAM tissue-derived growth factors of the same type, and when the mammalian tissue is implanted. such that the mammal becomes substantially enlarged, vascularized, and adapted as a result of Tissue is first implanted into or onto the CAM of a developing amniotic egg for a predetermined period of time. A composition that is