CN1218109A

CN1218109A - Production of organs for transplantation and method and apparatus for renovation and restoration

Info

Publication number: CN1218109A
Application number: CN 98119306
Authority: CN
Inventors: 任春严
Original assignee: Individual
Current assignee: Individual
Priority date: 1997-09-22
Filing date: 1998-09-11
Publication date: 1999-06-02

Abstract

A method for preparing the embryonic tissue and organ needed by transplantation features use of clone and transgenosis techniques to prepare the embryonic tissue and organ, which have more complete form and function and no exclusion after transplantation. The transplantation protector simulates the shape and structure of the tissue and organ and can protect the transplanted object after external culture and transplantation. With transgenosis method and placental shielding function, human embryo can continuously grow in other female animal body or bird's egg.

Description

Transplant production and organ reparation, reconstruction methods and the device of required organ

The invention belongs to a kind of embryo's material and methods for using them that is used to transplant with the clone technology manufacturing.

Present clone technology is that somatic nucleus is made it to stop to be transplanted in the unfertilized ovum after the division by hungry method.Embryonic organ's transplanting has the transplanting of the tissue such as thymus gland, pancreas, but require blood shape one to, and the source of organ is few, need to carry out pathological examination.The technology that the purpose of this invention is to provide usefulness clone bion is made the required tissue of desirable transplanting and the method for organ.

The manufacture method of graft is that to enter to make in the enucleated oocyte its growth with the nuclear transplantation that will accept the transplanter be that the clone technology of bion produces with accepting the identical embryo of transplanter's gene, but and directly transplanting after the tissue on the embryo and organ branch cut off, can transplant, can freezingly preserve through after cultivating.

The manufacture method of multipole graft is the embryo who produces a plurality of different differentiation degrees with embryo's cutting techniques and embryonic cell nuclear transfer technology.Embryo's cutting techniques is that the cleavage-cell of body early embryo is cut apart, and each cell all can develop into a complete biont.The embryonic cell nuclear transplantation is to use the nuclear transplantation of embryonic cell to produce clone body in ovum.Termination of pregnancy on different differentiation and development degree produces the embryo of a plurality of different differentiation degrees.

The manufacture method of totipotency graft is early stage the embryo, and termination of pregnancy when embryo's cell has totipotency is divided into several parts with embryonic cell.Can use transplanting protection device when in vitro culture and transplanting.Here the totipotency of mentioning is meant, but embryonic cell in being transplanted to body after differentiation and development become any cell of health, but become the totipotency difference of a complete biont with a cell differentiation and development during the embryo is cut apart.Totipotent transplanted thing generally is neural embryo and the embryo's material before it.

The manufacture method of versatility graft be embryo's neurula to organ differentiation to the entity of complete structure and function before, termination of pregnancy, with the embryo by differentiation and development after the difference of formed cell category cut apart, each class embryo material is divided into several parts.But this moment the cell on the embryo in migrating to body after the differentiation and development cell that becomes in natural embryo's differentiation and development, can differentiate.With embryo's material separately, various types of embryonic cells can not mixed in transplanting by different kinds.The absolutely most neurula stage the embryo of pluripotent cell exists in early days to fetus.

Manufacture method that specially can the property graft be foetal period organ the physiology and morphology function all form rear terminal pregnancy, each tissue and organ on the fetus are divided respectively and cut off, divide the tissue and the organ that cut off freezingly to preserve, can smash to pieces and make suspension, can cultivate in external continuation.

The cell in each differentiation and development of embryo period and tissue have different differentiation capabilities, activity and with the compatibility of accepting the transplanter.Transplanting to embryo's material all was to transplant with tissue and the organ of fetus in the past, ignored body early embryo and mid-term the embryo the advantage of the aspects such as differentiation capability, developmental potency and histocompatbility.Volume is more big in the transplanting of brain tissue and cell, and the more many graft of quantity is because the same oxygen supply of nutrition, and the effect after the transplanting is just more poor with respect to a small amount of transplanting.And with body early embryo and mid-term the embryo material transplant because embryonic cell can continue differentiation and development after transplanting, also can obtain good result so transplant a small amount of cell.

The advantage of totipotency graft is to have totipotent differentiation and development ability, extremely strong activity and histocompatibility, can transplant at any in vivo position.Its weak point is that the obtainable material of each embryo is few.The advantage of versatility graft is to have stronger differentiation capability, and extremely strong activity and histocompatbility can obtain more transplanting material.Specially the advantage of energy property graft is can obtain to transplant in a large number material and tissue and organ that certain physiology and morphology function is arranged.

The transplanting material of three kinds of etap can be mixed and use.When carrying out a large amount of cell transplantations, can sexual cell be main versatility or the totipotent cell of adding specially for example.Can be coated on surface and nervus vasculairs and the connecting portion with versatility or totipotent cell when organizing with the organ integral transplanting.The remodeling method of graft is to make the inborn defective that exists in the ovum of the embryo that cloned or normal fertilization obtain correcting by the specific gene that changes graft, make tissue and the organ of not complying with one's wishes on form or the function obtain desirable improvement, repulsion after the gene alteration that iuntercellular is identified is mutually avoided transplanting, can change differential growth speed, the aging rate of tissue and organ, the ability with impaired rear self-regeneration and regeneration is strengthened.Can change the tire phase before the clone changes by the gene of clone cell, after nuclear transplantation and after fertilization and change gene early stage embryonic stage.Can import other people or animal and plant genetic fragment, but artificial induction, manually synthetic or remove certain fragment gene.

The rejection that the heteroplastic transplantation of organ-tissue produces mainly is because of a fragment gene sequence that has iuntercellular mutually to identify in the gene, and in the past heteroplastic transplantation mainly is to alleviate rejection by medicine and chemicotherapy.The gene that this section iuntercellular of accepting the transplanter is identified mutually imports in the clone embryos or the brephoplastic graft that produces among the embryonated egg of normal fertilization or the cleavage-cell is implanted in this people's the body and will produce rejection.

In the human body more or less, either large or smallly all exist defective gene, what have reflects on the body of oneself, what have will just can reflect in the next generation or several generations.Can remove defective gene, insert normal foreign gene and can make natus more healthy, transplant with the organ of this fetus, the people of birth defect is rehabilitated.Importing has the gene of specific immunizing power, can obtain to have the immunocyte of extremely strong immunizing power.Hiv virus is because its characteristic apt to change is threatening human life always.Thereby eliminate hiv virus and can debate gene constant in the other hiv virus at the intravital immunocyte of only a few people.The genetic fragment of this section opposing AIDS virus is imported among the embryo, and the fetal immune system transplantation of generation enters in the AIDS human body can thoroughly eliminate AIDS.The cause of cancer is the somatic cell gene variation, produce the unrestricted auxocyte group who not controlled by self, and autoimmunity cell None-identified and remove these cancer cells, by importing foreign gene and can thoroughly curing cancer through the ability that the immunocyte that cancer cell activates increases fetal immune system identification cancer cell.

The gene fragment that inborn ex intelligence man is influenced IQ imports the embryo, again with the corresponding brain cell transplantation of embryo to the corresponding position of people's brain, the height of IQ depends on factors such as current potential that neuronic quality and quantity that the quantity of cell, cell are told and cell produce and quality of signals.To determine the embryo of IQ gene or human fetal brain or cell to be transplanted to memory and thinking district with the ex intelligence man, and can increase the quality of brain cell quantity and cell.Can make in the brain because of reasons such as ischemic, damage, agings by transplanting and to damage or regenerated in downright bad cell mass or a certain zone brain cell.Constantly carry out brain transplant with cerebral tissue aging, the life-span of brain is extended to infinitely impaired.

Tissue and the differentiation of organ, the speed of g and D depend on gene, and the change of the specific gene that control particular organization and Organ Differentiation are grown can make the differentiation of particular organization and organ, growth, aging rate change.Differentiation, speeding up of growth can make specific tissue and organ reach maturity as much as possible before terminal pregnancy, make it be suitable for transplanting.Produce the speed that contacts and mix with receptor tissue after changing transplanting, transplanted thing is grown as early as possible in vivo, improve transplantation effect.The development rate that slows down can be slowed down the aging of cell.

Foreign gene can be used the artificial synthesis manufacturing, can obtain from the body cell of other people with it, also can obtain from other biological cell.Animal is better than the mankind in the characteristic aspect of many heredity, and for example immunity system of Chelonian and the old and feeble gene of control can the regenerated genes after tissue of some animals or organ are impaired.The gene that imports the physiological change of cold-blooded animal or hibernator cell ischemic, anoxic and hibernation can make the easier preservation of graft and improve the transplanting quality.

The using method of graft is to the treatment process of graft before transplanting.

The integral transplanting method be with the upper effigurate tissue of embryo and fetus or organ integral body or transplanting partly before; available accept transplanter and fetus with it the body fluid of relevant position soak or flushing; with fetus have versatility or totipotent be divided into blood vessel and neural cell be coated in graft corresponding and accept transplanter's body on the nerve and blood vessel that link to each other, can be coated with versatility or totipotent nerve cell and vascular cell at the neural blood vessel pipeline of simulating when transplanting with transplanting protection device.Tissue and organ that the transplanting protector will be transplanted by the corresponding position are wrapped or fixed; each neural and blood vessel inserts on the transplanting protector in the mimic respective line; each junction of partly transplanting protector is sewed up; during transplanting can with body in each one of linking to each other be coated with tethelin and impel the protein molecule of cell attachment; as cold analysis sphaeroprotein or fine sticking plain, increase respectively connects two Jie's magnesium cations and the cationic content of two Jie's calcium.

Spreading method and injecting method are with graft and the distinctive body fluid growth hormone in position of accepting to transplant before transplanting.Impel the protein molecule of cell attachment to mix, increase the content of calcium magnesium cation.The spreading method spreads upon the position of accepting transplanting in transplanting, available sheet transplanting protection device covers, and prevents that cell runs off.Injecting method is with the transplanted cells of handling well the syringe of packing into, is expelled to accept the position of transplanting, and in tissue and the organ, cell can carry out microcarrier or microencapsulation is cultivated, the anti-mechanical shear stress of the activity of raising cell.But to infection such as the difficult position injecting immune cell treatment of arthritis bacterium that arrives of the immunocytes such as articular cavity and viruses.

Increase the growth hormone of graft, the body fluid that impels white matter and the calcium magnesium cation of cell attachment and accept transplantation site can impel behind the graft rapidly with body in corresponding tissue connect, and begin to work.In the pipeline of the analog neuron of transplanting protection device and blood vessel, be coated with to make with corresponding embryonic cell and extended to the connection that forms neural and blood vessel between the acceptor that can't contact and donor.

Transplanting protection device has internal structure and the external structure similar with organ to transplanted tissue; it is several partly independent to be used for protecting transplanted tissue and organ, transplanting protection device to divide, and partly the junction stitching just can be with complete the wrapping of transplanted thing with each after graft can being wrapped from each side.

The material of transplanting protector has biological degradability; and contain in hormone embryo that promotion grafts such as aerobic, tethelin, adherent hormone grow in vivo and the adult nutritive substance contained in the body fluid of corresponding position and ion and in transplanting the process that protector is degraded in vivo discharging gradually, promote to transplant the growth of material.

It is that fetus or the corresponding brain cell transplantation of embryo are gone into corresponding position in the brain that embryo and brain is transplanted the method that improves IQ, by improving the structure of brain, changes the gene of brain cell IQ and the gene of personality, and brain is improved on the amount of mass of cell.Brain cell can be transplanted to IC memory district, and memory can improve thinking ability to thinking zone transplanting, transplants the contact and the cooperation that can strengthen left and right sides brain in the left and right sides brain part that is connected.Improve the structure of brain with the method and can develop the brain that the end is utilized.

People's IQ is subjected to influences congenital and two factors day after tomorrow, the quantity of decision brain cell, the neuronic quality and quantity that each brain cell is told, the ability of cell acceptance and output information.

Inborn factor is exactly the factor of gene, by can obviously improving embryo's congenital IQ to the change of early stage cleavage-cell's gene or the change of normal fertilization ovum gene behind the change of nuclear transplantation nucleus intelligence gene, the nuclear transplantation, also available this method improves newborn baby's IQ.Acquired disposition be fetus in parent, the sign stimulus of natus and underage child brain impels brain cell development.The brain cell transplantation of accepting the embryo of ex intelligence man's genetic modification or fetus can be significantly improved people's IQ in adult's brain.Even the fetal brain cell that does not have ex intelligence man's intelligence gene is in the IQ that also can significantly improve the people after the transplanting owing to the stimulation that is subjected to external information on quality and quantity.

The method that heterogenous animal breeds the embryo is to impel animal embryo to grow in heterozoic parent or in the ovum by gene that changes the embryo and the parent of becoming pregnant and the environment of simulating both sides' implantation.

Because idea and the impact of miscarriage to human body of becoming pregnant, the embryo and the fetus that supply transplanting usefulness that the ovum of employment and people's parent manufacturing is a large amount of are not-so-practical.Because the embryo has certain flexibility in the immunity of transplanting, embryo's trophocyte does not express Human leucocyte antigen-Ⅰ class antigen, and fetus is not repelled by parent.And embryo's immunity system does not form, and the complete antibody that is beyond expression makes the embryo be easier to be accepted by heterozoic parent.By changing specific gene or temporary gene expression, can change the expression of the antibody of fetus or parent, further avoid the generation of rejection.Can remove immunogene or xenogenesis maternal immunity gene, or make parent produce the transplantation immunity tolerance, the chimeric technology human embryos of available embryo are transplanted to other Mammals and are removed embryo's main body, only on the embryo of remaining placenta.

The implantation mechanism that can change fetus by the specific gene that changes the embryo is namely changed into embedding implantation or surperficial implantation by original complete invasive implantation.Also can change the parent form of becoming pregnant by changing the parent specific gene.By changing specific open gene nucleus can be grown in the heterogenous animal ovum, and can examine to realize by transplanted cells in the heterozoic ovum of the stoning of being fertilized.

Change specific gene, mammiferous embryo is grown in the ovum of birds.Mammals is very similar when the differentiation and development with the birds embryo, and just birds are based on the yolk sac circulation, and especially human yolk sac and its circulation of Mammals all degenerated, and the substitute is placenta.Its embryo's cleavage rate and rate of development also than birds slowly many.Birds yolk sac circulation development gene is replaced the placenta development gene, improve fetal development speed and can make Mammals in birds eggs, set up the recycle system.

Mammiferous embryo to birds eggs transplant can be divided into transplant in the body and external ovum in transplant.Implantation method is to choose the ovum of foundation fully but yolk sac circulates not unfertilized or that be fertilized in the body, ovum is taken out, embryo in the ovum is taken out, implant mammiferous embryo, egg membrane with same fertilization or unfertilized egg takes off some, with bonding or sew up and to remedy the egg membrane breakage that causes because of transplanting, again with fertilization place of ovum transplantation ovum in the parent.The egg that produces by chicken have be fertilized can prove that with unfertilized phenomenon the resolving ability whether birds be fertilized to ovum is very poor.The ovum that the artificial graft enters can equally with other normal fertilization ovum be wrapped outer egg membranes such as albumen eggshell, comes out under normal ovum.Because the ovum of transplanting has removed chorion, and is very weak, so will there be thing when transplanting, to protect.Shown in Figure 1 is the protector that supplies ovum transfer." 1 " is two the same semicircle things of spoon among Fig. 1, can be transplanted ovum protection interior after the merging. " 2 " are to close backguy, link to each other with " spoon ", make two " spoons " as scissors." 3 " are to decontrol backguy, and during the line tension, two " spoons " branch is opened to ovum." 4 " are the axles that connects two " spoons " and center rod.Can emit water polo " 5 " between two spoonfuls ovum is ejected then two spoonfuls of merging.Water polo is on center rod " 6 ", and center rod hollow can push water polo in external water filling, and water polo expands, and ejects ovum.Water polo should have very snappiness, can not break.The chimeric implantation technique of available embryo carries out in the body or external transplanting.Mammal can be transplanted in the birds embryo or remove in the birds embryo who only stays the circulatory system of main body, grow by the circulation of birds.

External transplanting is the zygote or the unfertilized egg period that go out under ovum before back to vitelline vessel is set up, and the chorion and the tunica fibrosa of embryo present position are scratched, and stretches into the semicircle arc clip and clamps ovum, and the ovum frenulum can not magnetic breaks.Plasma membrane and vitellinae membrana are scratched, taken out intravitelline embryo, implant mammiferous embryo, plasma membrane is sewed up with vitellinae membrana or the bonding or stitching with film identical in same fertilization or the unfertilized ovum.Tunica fibrosa in chorion and the shell also can be mended like this and close.The semicircle arcuation clip that the folder ovum is used can merge with clamp holder after the radian of chorion incision stretches into chorion and clamp ovum earlier in two separately again.The ovum frenulum of ovum can stretch into clip from the centre only at the two ends of ovum.

After the embryo of the fetus that needs long term growth is placed in the ovum, can be with the ovum portion support device at chorion cutting section oxygen gas-supplying, nutriment and moisture content.The moisture content that the embryo consumes in growth and nutriment derive from the white of an egg and yolk, and oxygen exchanges from air by eggshell.The fetal growth time is much longer with respect to the chicken embryo, and moisture content evaporates from eggshell easily.Get radian eggshell similar to the place of being cut open and together cut a hole together with tunica fibrosa, the keeping mouth of ovum portion support device is placed on the hole of incision, and protection sealing around making.Eggshell is placed on the incision of the ovum of accepting embryo transfer together with the ovum apparatus of oxygen supply, two eggshells are bonding.The part at apparatus of oxygen supply and the white of an egg place of connecing can arrange a suction inlet, and an outlet, suction inlet are with egg white sucking-off from ovum, and row's mouth will add moisture content, and the egg white of nutriment and oxygen flows back in the ovum.Can be at the same place of cutting of the opposite of chorion cutting section chorion, one of two cutting section are laid suction port, lay relief outlet for one, and this method also helps the detection of fetal development and observation.Can be easy to observe embryonic development at peephole of ovum apparatus of oxygen supply placement and lamp.

Ovum can be put into culture medium or remove eggshell and put into culture medium, can draw the nutrition in the culture medium.And can be with the cutting of small emery wheel when removing eggshell, the distance that the emery wheel edge exposes backplate is not more than the thickness of eggshell, prevents to injure the ovum under the eggshell.Also the available enzyme digestion method impels cytolemma to separate with chorion.After the embryo sucks lecithocoel with the white of an egg, tunica fibrosa can be cut, make that material in the culture medium is easier to be exchanged with the embryo.

The essentially consist material of various zooblasts is big to identical, but the demand of nutrition is had nothing in common with each other.Method with temporary genetic expression when embryo transfer conforms the embryo in a short time, can in parent, inject the progestogen that is produced when parent is become pregnant, the preparation that parent is carried out become pregnant, to the secretory product of the intravital embryo's embryonal vaccination of mother parent excretory of the same race uterine milk, the embryo and the parent of becoming pregnant are become pregnant under more satisfactory condition with the embryo of the parent preparation implantation of the same race of becoming pregnant.And when transplanting mammal embryo in the birds eggs, nutriment and immunizing antigen and the immunoglobulin (Ig) that can lack to uterine milk and the birds of the parent secretion of the same race of embryonal vaccination in yolk and the white of an egg together with the embryo.Diet regulation and control to parent can solve parent and the difference of embryo on nutritional need to a certain extent, can be by inject the nutriment that replenishes the embryo in parent and in ovum.

The advantage that heterogenous animal breeds the embryo is to make the embryo of rare animal to find more parent, and human inconvenience and the injury that can break away from conceived miscarriage.Human embryos are bred in other mammal or birds eggs, make the easier acquisition of embryo of human each differentiation and development phase, for producing transplant organ and helpful to the further investigation of embryonic development.The specific gene that change is examined by clone cell, the specific gene of zygote and the specific gene of body early embryo can make human somatic cell nuclear, the nucleus of zygote and embryo's nuclear transplantation go in the stoning ovum of heterogenous animal and birds, in the early stage cleavage-cell of stoning, make the embryo continue to grow adult's embryo.Can make human the minimizing rely on human ovum to make the embryonic organ who transplants usefulness.

It shown in the Figure of description 1 protector for ovum transfer.Among the figure " 1 " be two semicircles as spoon, be combined and protect in the middle of can being combined in the ovum of the birds of removing chorion." 2 " are to close backguy, and " 4 " are axis, are " spoons " and the cutting apart of following " spoon ", close backguy " 2 " tension after, be axle with " 4 ", the stressed stressed closure when the scissors closure of two " spoons "." 3 " are to decontrol backguy, link to each other with " the spoon back of the body ", and " the spoon back of the body " is stressed after the tension, and two " spoons " open." 5 " are water polo, and " 6 " are center rod.Center rod " 6 " hollow, the top connects water polo " 5 ", can water filling in water polo " 5 ".Institute's water filling is nutrient solution, and both having made breaks can not injure birds yet.Water polo " 5 " ejects ovum through dilation by water injection, two spoonfuls of closures.

Be that mammal nuclear or embryo, zygote transplation are clamped ovum in the birds shell with clip in the birds eggs shown in the Figure of description 2.Be the spoon shape clip of two semi arches shown in " 1 " among the figure, the clip side open up less than 60 °, allows the ovum frenulum of ovum both sides, and circular open is arranged on the clip, can abdicate the birds embryo and grasp for the behaviour person of being and do.Opening on the chorion is the smaller the better, so two " spoons " are put into ovum along the radian of ovum respectively, in the slot " 4 " of kettleholder " 2 " insertion " spoon " frame " 3 ", just can be used for clamping ovum.Spring piece is used for controlling the dynamics of ovum.Spoon can be provided with a plurality of minimum sons that suck in the class.Suck the son back and connect little vacuum-lines.When clip is clamped ovum, make by little vacuum line and suck son generation vacuum, the fixedly ovum even clip is not exerted oneself also.Be communicated with the external world in the complete final vacuum pipe of graft procedure, vacuum will disappear.Can in the white of an egg, support by valve tube.

The purpose of manual simulation's growing environment device provides in the similar parent and the intravital back of people dis environment, composition, various trace element, ion and isoionic environment, intravital bioelectricity electric current, electric wave, hertzian wave, electric field and the magnetic field of simulation nutrition are made up of measuring apparatus, regulation device and manufacturing installation.The equipment that equipment such as electrocardiograph, electroence phalograph, electromyograph, stomach and intestine electrograph machine and physiatrics are arranged in the market.Before graft is transplanted with its nerve, blood vessel with body in the position that is connected connect corresponding bioelectrical signals and simulated blood vessel, in substratum, add materials such as electrical conductor, magnetizer and add corresponding electric field, magnetic field at the blood vessel two ends, and can be by the growing environment of blood vessel and neuromodulation graft.

The key step of manufacturing when transplanting of clump graft shown in the Figure of description 3.Among the figure " graft " refer to comprise behind the clone, normal fertilization, through the material of transplanting of the ovum of genetic modification and cleavage-cell, embryo, fetus and need through manual simulation's growing environment cultivate, in the human parent, in the animal parent and the material that continued growth is grown in the animal ovum.Gather cell on can the clump graft and carry out clone and the genetic modification of embryonic cell.

Claims

One kind with the cloning process manufacturing with accept transplanter's gene one to tissue and organ and can make its form and the better method of function, form by the manufacture method of graft, the remodeling method of graft, the using method of graft, method, transplanting protector and the manual simulation's growing environment device that heterogenous animal breeds the embryo, the manufacture method that it is characterized in that graft is that the nucleus that will accept the transplanter is made the embryo with the technology of clone biont, and embryo's material is used for transplanting; The remodeling method of graft is the specific gene that changes transplanted thing, reaches the form and the function that need; The using method of graft is the method for handling and using of giving before transplanting; The method that heterogenous animal breeds the embryo is that the gene that changes the embryo and the parent of becoming pregnant makes a kind of animal can borrow ovum of another kind of animal and parent to grow with the condition of simulating both sides' implantation; Transplant protector by depending on growth for cell, permission has permeability to small-molecule substance, contain conductor material and different kinds of ions material, but there is the material of certain elasticity and degradation in vivo to make, in material, can add oxygen, tethelin and nutritive substance, in the transplanting protector is degraded process, discharge; Manual simulation's growing environment device is the metering facility and the device of making the ionic environment that measures by various ionic environments.
2. according to claim 1, the manufacture method that it is characterized in that graft is in the egg cell of the nuclear transplantation that will accept the transplanter end fertilization that enters stoning, make it develop into the embryo, organ on the embryo can be used for transplanting, but with embryo's material by the tissue and organ cut apart after directly transplanting can freezingly preserve, also can continue to cultivate rear transplanting; The manufacture method of multistage graft can be made Gong the tissue in a plurality of different differentiation and development stages and the embryo that organ transplant is used.
3. according to claim 2, the manufacture method that it is characterized in that multistage graft is the embryo to be carried out the embryo is cut apart or the embryonic cell nuclear transplantation, produce the embryo of a plurality of different differential periods, the embryo who produces is cut apart by the characteristic of different differential periods, according to the differential period difference, can be divided into totipotency and transplant the manufacture method of material, versatility is transplanted the manufacture method of material and the manufacture method of special energy property transplanting material;

The manufacture method of the transplanted material of totipotency is when embryo differential period in early days, any one cell of embryo all can be divided into the stage of any cell of any tissue of whole body and organ after transplanting, to cut apart the totipotency that can develop into another complete biont incomplete same with the embryo, this moment, termination of pregnancy was divided into several parts with embryo's cell; The manufacture method of the transplanted thing of versatility be the embryo break up reached maturity to the form of organ and physiological function before, a kind of cell also becomes differentiation and development various kinds of cell period, the embryo is cut apart, cutting apart of existing certain morphological structure by form integral body, certain form is not arranged, different separated by the cell category that can break up thereafter, every kind of cell is divided into several parts; Specially the manufacture method of the transplanted thing of energy property is in embryo tissues and organ certain form and physiological function to be arranged, the well differentiated special energy sexual stage of cell, and stop appointing and be pregnent, the embryo is cut off by different tissues and organ branch.
4. according to claim 1, the remodeling method that it is characterized in that graft is by the gene that changes the embryo the specific tissue of embryo and organ to be improved at morphology and function; Remodeling method can be to import his people's gene or animal-plant gene or cause the autogene sudden change or remove specific gene; Change the gene of cell before the nucleus of transplanting takes out from cell, culture expression is picked out the normal cell of transgenosis and is carried out nuclear transfer then; Can behind nuclear transplantation, to spilting of an egg knot end, carry out genetic modification, can carry out genetic modification to zygote cell and the cleavage-cell of normal fertilization; Can behind the embryonic cell quiding gene, carry out nuclear transfer; Can be when embryo separating quiding gene;

The manufacture method of not having the rejection brephoplastic graft is to import the genetic fragment of accepting the produced immune response that cell is identified mutually in transplanter's gene in fertilization with in the ovum after cloning and in the quilt cell of cloning, the antigen that the antigen that the graft that produces is transplanted rear generation can not intensify the immune system of accepting the transplanter and be accepted the transplanter intensifies, immunocyte and the anti-source of accepting the transplanter are expelled among the embryo, can make the compatibility of embryo's identification and formation docking host's antigen;

The graft function Innovative method is by changing specific gene, specific tissue and organ are improved at morphology and function, transplant with the brephoplastic graft after improving, can make original change that has the tissue that adds earlier defective and organ to obtain essence, change gene to embryonated egg and can produce muscle power, intelligence, life-span and resist the better fetus of the original gene of disease geometric ratio;

The method of increase and decrease allelotaxis speed is: by differentiation and development speed and the old and feeble gene that changes particular organization and organ, accelerate the speed of the interior certain organs of embryo and tissue, make the embryo can be competent at a job after transplanting in this certain organs of fetal period, but and delaying senility.
5. according to claim 1, the using method that it is characterized in that graft is to cultivate the back with sugar degree and the higher nutrient solution of oxygen level to be coated on the transplanted thing with the distinctive body fluid of accepting to transplant in position before transplanting, when transplanting, be coated with tethelin, and adjust various ionic content, can be divided into integral transplanting method, spreading method and injecting method;

The integral transplanting method is before transplanting, in culture medium, cultivate or directly transplanting with transplanting protection device, in the simulated blood vessel of transplanting protection device, the nerve channel with body in the junction be coated with the corresponding nerve of formation, blood vessel and the cell that is connected on the embryo; The spreading method be with embryo and fetus applying materials at the position of accepting to transplant, and lid prevents that with the sheet transplanting protection device cell runs off; Injecting method is that embryo and fetus material are put into syringe, is expelled in the position and histoorgan of accepting in the body to transplant; When transplanted thing can't work alone, formerly in the body can keep with organ in a organized way, treat the rear excision that can work independently of transplanted thing; Former in a organized way with keeping in vivo that organ still can be survived.
6. according to claim 4 and 5, it is characterized in that method that embryo and brain transplant to improve brain function is that embryo or the corresponding brain cell of fetus or tissue transplantation are gone into corresponding position in the brain; Transplant can be without genetic modification or the receptor gene transformation, import the brain material of ex intelligence man's IQ gene fragment, the brain function of accepting transplanter and baby is improved under the raising of cell quality and quantity.
7. according to claim 1 and 4, it is characterized in that method that heterogenous animal breeds the embryo is both sides to be accepted mutually and the principle of simulation both sides implantation can promote generation and the success of implantation by the gene that changes embryo and the parent of becoming pregnant;

Gene alteration is can change the repulsion of become pregnant parent and embryogenesis and the growth of implantation mechanism and placenta by genetic fragments such as the immunity identification gene that imports the other side, placenta, yolk bag circulation development genes, change hormone and the nutrition of become pregnant time and secretion, can change embryo's rate of development;

Implantation principle simulation reconstruction is that the secretory substance before and after implantation takes place is expelled in the parent body with parent and its zygote of the same race, stimulating parent to carry out implantation prepares, and the parent that the embryo is of the same race and the zygote secretory product before and after implantation is expelled to the implantation position, nutrition is provided and stimulates embryo nidation;

Mammal borrows birds eggs to come embryonic development by strengthening the function of embryo's yolk bag blood circulation system, to accelerate the growth of blood circulation and increase corresponding hormone, growth factor and nutriment in ovum.
8. according to claim 7, it is characterized in that cloning with oviparous animal is the oviparous animal that produces with the genetic modification method, the ovum that adults is given birth to can not have nucleus, but differentiation and development becomes the embryo after implanting the nucleus of being cloned; Give birth to and can contain nucleus in the ovum, restrain after taking out ootid nucleus; Grand; Spermatogonium, the sperm mother cell of accepting clone's species can be implanted in the oviparous animal parent; Can make the state of activation that is in after the egg cell maturation before the spilting of an egg with the method that changes gene, can make the nuclear gene in the ovum not produce genetic expression, can automatically clear up, when the clone, it need not be taken out.
9. according to claim 1, it is characterized in that transplanting protection device is by depending on growth for cell, permission has permeability to small-molecule substance, but there is the material of certain elasticity and degradation in vivo to make, in material, can add oxygen, growth hormone and nutriment, in transplanting protection device is degraded process, discharge; The connection that when the each several part of transplanting protection device all is distributed with when the transplanted thing growth of impact, easily is bursting at the collision; Shape and structure after the shape of this device and the constructing analog graft growth and maturity have blood vessel, the nerve of simulation and are connected the structure that connects; Can be decomposed into some parts, be divided into each orientation of several part clumps according to the structure of graft and encase or support transplanted thing, form an integral body after merging, each partly between available stitching or bonding method connect; The material of available this device is made the arbitrary shapes such as sheet, bar-shaped, tubulose; plant protection protection unit and graft can be made the characteristic that contains the described transplanting protection device of claim 8 with the micronize in the cell culture technology, microencapsulation, embedding method, covalency attaching method, ion covalent cross-linking method, can adapt to local transplantation, nerve, blood vessel, bone and cell the requirement of transplanting.

Can be divided into packing shape, carry shape and mix shape according to the structure difference; The external structure of packing shape is similar to the external structure of graft, encases graft in culture apparatus and in the body during transplanting; The internal structure of carrying the structure of shape and transplanted thing is similar, makes transplanted thing can be attached to top growth, can support transplanted thing growth; Mix shape and have packing shape and carry the shape feature, can clump inner support and the transplanted thing of clump outer protection.
10. according to claim 1 spouse, it is characterized in that manual simulation's growing environment device is by the measuring apparatus that comprises various ions, plasma ambient and electric microfield, micronormal and can makes the substratum of the all-environment device of measuring gained and by the artery that nutrient solution can be pumped into tissue and organ by the frequency and the dynamics of pulse, and the clump vein is extracted the pump composition of nutrient solution out.