Bank of stem cells for producing cells for transplantation having HLA antigens matching those of transplant recipients, and methods for making and using such a stem cell bank
View Patent ↗Methods for producing stem cell banks, preferably human, which optionally may be transgenic, e.g., comprised of homozygous MHC allele cell lines are provided. These cells are produced preferably from parthenogenic, IVF, or same-species or cross-species nuclear transfer embryos or by dedifferentiation of somatic cells by cytoplasm transfer. Methods for using these stem cell banks for producing stem and differentiated cells for therapy, especially acute therapies, and for screening for drugs for disease treatment are also provided.
1. A library comprising a plurality of isolated, genetically modified human embryonic stem cell lines produced by the method comprising:
(a) genetically modifying a checkpoint arrested, senescent or near senescent human donor cell to be homozygous for at least one MHC allele by the removal of one or more sets of MHC alleles by knockout or RNA interference or the addition of another homozygous set of MHC alleles;
(b) isolating the nucleus from the genetically modified donor cell;
(c) inserting the nucleus into an enucleated human oocyte to produce a nuclear transfer embryo;
(d) activating said nuclear transfer embryo and culture the embryo until it forms a blastocyst comprising a discernable inner cell mass; and
(e) isolating cells from the inner cell mass to produce an embryonic stem cell line
wherein each of said plurality of stem cell lines is homozygous for a different set of MHC alleles relative to the remaining members of the plurality of stem cell lines, wherein the stem cells have EPC-1 and telomerase activity greater than EPC-1 and telomerase activity in age-matched control cells of the same type that are not generated by nuclear transfer techniques.
2. The library of claim 1 which is comprised of at least five different stem cell lines, each member homozygous for a different combination of histocompatibility or MHC antigen alleles.
3. The library of claim 1 which is comprised of at least ten different stem lines, each member homozygous for a different combination of histocompatibility or MHC antigen alleles.
4. The library of claim 1 which is comprised of at least about 100 to 1000 different stem cell lines, each homozygous for a different combination of histocompatibility or MHC antigen alleles.
5. The library of claim 1 wherein all of the stem cell lines are O-negative.
6. The library of claim 1 wherein the library includes human stem cells that are homozygous for one or more of the following MHC alleles: HLA-A1, HLA-A2, HLA-A3, HLA-A24, HLA-A11, HLA-A28, HLA-A29, HLA-A32, HLA-B15, HLA-B5, HLA-B7, HLA-B8, HLA-B12, HLA-B14, HLA-B18, HLA-B35, and HLA-B40.
7. The library of claim 6 wherein all of said human stem cell lines are O-negative.
8. The library of claim 1 which includes stem cell lines which are homozygous for at least one of the following HLA-A, HLA-B and HLA-DR haplotype combinations: 1, 7, 2; 1, 8, 3; 2, 14, 1; 2, 35, 4; 2, 35, 8; 2, 44, 4; 3, 7, 2; 3, 7, 4; 3, 7, 8; 3, 35, 3.7, 8; 3, 35, 1; 31, 51, 4; and 32, 14, 7.
9. The library of claim 8 which contains cell lines homozygous for all of said HLA-A, HLA-B and HLA-DR haplotype combinations.
10. The library of claim 8 which is O-negative.
11. The library of claim 9 which is O-negative.
12. The library of claim 1 , which contains a stem cell line homozygous for an MHC, allele selected from HLA-A, HLA-8, HLA-C, HLA-DR, HLA-DQ, and HLA-DP, wherein the cells are of the O-negative blood group.
13. The library of claim 1 , which contains a stem cell line homozygous for the MHC alleles encoding HLA-A, HLA-B, and HLA-DR.
14. The library of claim 1 , which contains a stem cell line homozygous for the MHC alleles encoding HLA-A, HLA-B, and HLA-DR, wherein the cells are of the o-negative blood group.
15. The library of claim 1 , which contains lines of diploid stem cells in which all of the MHC alleles are homozygous.
16. The library of claim 1 , wherein the stem cells have telomeres that are on average at least as long as the telomeres of age-matched control cells of the same type that are not generated by nuclear transfer techniques.
17. The library of claim 16 , wherein the proliferative life-span of the stem cells is at least as long as the proliferative life-span of age-matched control cells of the same type that are not generated by nuclear transfer techniques.
18. The library of claim 16 , wherein the proliferative life-age-matched control cells of the same type that are not generated by nuclear transfer techniques.