CN102286427A - Method for producing hematopoietic stem cells of human being with animals - Google Patents
Method for producing hematopoietic stem cells of human being with animals Download PDFInfo
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- CN102286427A CN102286427A CN2011102552243A CN201110255224A CN102286427A CN 102286427 A CN102286427 A CN 102286427A CN 2011102552243 A CN2011102552243 A CN 2011102552243A CN 201110255224 A CN201110255224 A CN 201110255224A CN 102286427 A CN102286427 A CN 102286427A
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Abstract
The invention belongs to the field of bioengineering, and particularly relates to a method for producing hematopoietic stem cells of the human being with animals. The method includes the following steps: (1) the somatic cell of a patient is used for producing an embryonic stem cell (ES) or an induced pluripotent stem cell (ips); (2) the produced ips or ES is injected into the blastula of an animal, and the blastula is then transplanted into the pseudopregnant animal uterus, and is developed into a 'human-animal chimera' embryo; (3) the hematopoietic stem cells of the human being are separated from the bone marrow, placenta and umbilical cord blood of a delivered 'human-animal chimera' fetus by the immunomagnetic bead separation method. The method has the advantages that: the source of hematopoietic stem cells is abundant, the genetic backgrounds of the hematopoietic stem cells and the patient are alike, the age of the hematopoietic stem cells is infant, and therefore the proliferation and differentiation potentials of the hematopoietic stem cells are higher than the proliferation and differentiation potentials of the hematopoietic stem cells from an adult.
Description
Technical field
The invention belongs to bioengineering field, particularly relate to the hemopoietic stem cell of producing the people with animal.
Background technology
Hemopoietic stem cell can be divided into multiple histocyte, disease such as heart trouble, leukemia can obtain medical treatment by transplanting hemopoietic stem cell, but the genetic background of used hemopoietic stem cell must be close with the patient or identical, otherwise can suffer patient's immunological rejection, but, be difficult to the donor that finds genetic background similar, and find the identical donor of genetic background hardly may to the patient.In the general population, find only hundreds of thousands of/one of the similar donor of genetic background, just as looking for a needle in a haystack.Therefore, a lot of patients finally still can not find the ideal donor and have unfortunately left this world.Even found the similar donor of genetic background, because gene does not have identically, the patient may also immunological rejection to a certain degree can occur and cause the not good enough or graft failure of curative effect the hemopoietic stem cell of donor.
The Britain scientist successfully people's hematopoietic stem cell transplantation in the body of pig and sheep embryo (being intrauterine transplantation), allow hemopoietic stem cell increase in animal body, the intravital human hematopoietic stem cell of separating animal's can be transplanted in the human body and play a role then; 2007, the embryo that the bone marrow stem cell of the scientist of Nevada ,Usa university personnel selection injects sheep still, the sheep body contains people's cell of 15%.But; the human hematopoietic stem cell that this method is produced; its age is the same with the people that hemopoietic stem cell is provided; add hemopoietic stem cell division growth in animal body; telomere can be shorter, and therefore, the human hematopoietic stem cell that this method is produced is also older and more feeble than the people that human hematopoietic stem cell is provided; also not as young hemopoietic stem cell, result of treatment is not satisfactory for its proliferation potential and differentiation potential.
Though embryonic stem cell (ES) or induced multi-potent stem cells (ips) can be divided into hemopoietic stem cell in vivo and in vitro, and the age of cell is the same with fetus inmature, but, directly ES or ips implant into body: (1) produces immunological tolerance phase cell before because ES or ips are the embryos, some antigens are arranged not through immunity identification tolerance, therefore, even genetic background is identical with the patient, the intravital immunity system of patient also can repel it as foreign matter, " Reference News " reports and is entitled as " artificial animal stem cell the failure of an experiment " 2011.5.16 day the 7th edition, say: the Britain scientist is transplanted to iPS in the mouse body and is excluded, cause my once (Beijing medical science communication, 1998 of this consequence; 66:1058,1060, house journal will be read this paper and need arrive Beijing, Beijing and teach by correspondence medical college and ask for) said that the stem cell that transforms (rejuvenation) may be expressed the embryo antigen that tolerance is not discerned in the fetus immunity, can cause immunological rejection; (2) can produce teratoma; (3) though ES under the condition of vitro culture or ips also can be divided into hemopoietic stem cell, but, per 3 cell fission once just have 1 sudden change to take place under the condition of vitro culture, and its sudden change has storage effect, therefore, after the process propagation of some generations, the most of cell in a group cell all suddenlys change, and does not have using value.
Life does not have the several fatal defective of above-mentioned human intervention generation usually at the embryo development procedure of nature.
Is 129-S mouse teratocarcinoma cell, being transplanted to C57-b is in the blastaea of mouse, then, again blastaea is transplanted to the intrauterine of other foster mother mouse, make its further growth, the result has given birth to and planted is that mutual miscellaneous is the normal allophenic mice of inlaying style, and all are organized, organ all is made up of the somatocyte of these two kinds of mouse (Mintz, 1978).
Summary of the invention
The objective of the invention is to produce people's hemopoietic stem cell, make the patient not need earnestly to seek and just can obtain the complete and own identical hemopoietic stem cell of genetic background with animal.
Produce people's hemopoietic stem cell by what this purpose provided with animal, its steps characteristic comprises:
1, embryonic stem cell (ES) or the induced multi-potent stem cells made from patient's self somatocyte (ips).
2, the ips that manufactures or ES are injected in the blastaea of animal, blastaea is transplanted in the animal uterus of false pregnancy and is developed into " people-animal mosaic " embryo then.
3, the people-animal mosaic that utilizes branch to give birth to is isolated people's hemopoietic stem cell.
The present invention has hemopoietic stem cell source abundance, and the genetic background of hemopoietic stem cell is identical with the patient, and the age of hemopoietic stem cell is inmature, and therefore, propagation is recently big from adult hemopoietic stem cell with differentiation potential.
Embodiment
Below the present invention is done and describe in further detail.
Produce people's hemopoietic stem cell by what this purpose provided with animal, its steps characteristic comprises:
1, embryonic stem cell (ES) or the induced multi-potent stem cells made from patient's self somatocyte (ips).
2, the ips that manufactures or ES are injected in the blastaea of animal, blastaea is transplanted in the animal uterus of false pregnancy and is developed into " people-animal mosaic " embryo then.Used animal blastaea can be pig, sheep or primates, and is better with pig, because the period ratio of pig growth and breeding is shorter, build is bigger.
3, the chimeric fetus marrow of people-animal, placenta, the Cord blood that utilizes branch to give birth to isolated people's hemopoietic stem cell with the immunomagnetic beads separation method.
The chimeric individuality of people-animal contains the cell of 4 types: 1, animal adult stem cell; 2, the non-stem cell of animal tissues; 3, people's adult stem cell is as hemopoietic stem cell, mescenchymal stem cell etc.; 4, the people organizes non-stem cell.
In general, the chimeric fetus of people-animal is difficult to survival in minute puerperium, if divide the chimeric fetus of people-animal of giving birth to survive, can raise the hemopoietic stem cell of certain period separation of human from marrow, or allow stem cell be discharged in the blood circulation with the stem cell mobilization agent, separate peripheral hematopoietic stem cells with whizzer then, improve the output of hemopoietic stem cell with this; The childbirth of not free animal by the time can be cut open the belly and be taken out the embryo ahead of time if the patient is badly in need of hemopoietic stem cell.In addition, also can from people-animal mosaic, isolate other adult stem cell of people, as mescenchymal stem cell.
In the technological line of the present invention, it all is ready-made making technology such as embryonic stem cell (ES) or induced multi-potent stem cells (ips), the transplanting of animal blastaea, hemopoietic stem cell separation with somatocyte, need not here to give unnecessary details, and please check pertinent literature.
Claims (1)
1. produce people's hemopoietic stem cell with animal, its steps characteristic comprises: 1, embryonic stem cell (ES) or the induced multi-potent stem cells made from patient's self somatocyte (ips); 2, the ips that manufactures or ES are injected in the blastaea of animal, blastaea is transplanted in the animal uterus of false pregnancy and is developed into " people-animal mosaic " embryo then; 3, the chimeric fetus marrow of people-animal, placenta, the Cord blood that utilizes branch to give birth to isolated people's hemopoietic stem cell with the immunomagnetic beads separation method.
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| CN2011102552243A CN102286427A (en) | 2011-08-28 | 2011-08-28 | Method for producing hematopoietic stem cells of human being with animals |
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| CN2011102552243A CN102286427A (en) | 2011-08-28 | 2011-08-28 | Method for producing hematopoietic stem cells of human being with animals |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107920499A (en) * | 2015-06-22 | 2018-04-17 | 全国农业协同组合连合会 | The production method of blood chimerism animal |
| CN111315881A (en) * | 2017-11-17 | 2020-06-19 | 国立大学法人筑波大学 | Non-human animals and methods of production thereof |
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2011
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107920499A (en) * | 2015-06-22 | 2018-04-17 | 全国农业协同组合连合会 | The production method of blood chimerism animal |
| US11432537B2 (en) | 2015-06-22 | 2022-09-06 | The University Of Tokyo | Method for producing blood chimeric animal |
| CN111315881A (en) * | 2017-11-17 | 2020-06-19 | 国立大学法人筑波大学 | Non-human animals and methods of production thereof |
| CN111315881B (en) * | 2017-11-17 | 2023-10-10 | 国立大学法人筑波大学 | Non-human animal and method for producing the same |
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Application publication date: 20111221 |