US7030292B2
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Method for producing a population of homozygous stem cells having a pre-selected immunotype and/or genotype, cells suitable for transplant derived therefrom, and materials and methods using same
Abstract A method of producing a homogenous population of homozygous stem (HS) cells pre-selected for immunotype and/or genotype from donor cells is described herein. The invention relates to methods of using immunohistocompatible HS cells for diagnosis, therapeutic and cosmetic transplantation, and the treatment of various genetic diseases, neurodegenerative diseases, traumatic injuries and cancer. The invention further relates to methods for using histocompatible HS stem cells pre-selected for a non-disease genotype for prophylactic and therapeutic intervention including, but not limited to, therapeutic and cosmetic transplantation, and the treatment of various genetic diseases, neurodegenerative diseases, and cancer. Furthermore, the invention relates to a catalogued transplant depository of HS cells derived from multiple donors, each of the HS cells being homozygous for a unique HLA haplotype, for the purpose of having a constant, reliable, comprehensive supply of immunohistocompatible cells for diagnosis, treatment and/or transplantation.
A method for producing a mammalian homozygous embryonic stem (HES) cell depository, wherein each HES cell produces a cell line, which has alleles that are homozygous for a Major Histocompatibility Complex (MHC) haplotype, wherein the method comprises: (a) selecting donors; (b) determining the MHC haplotype of each donor; (c) mitotically activating non-fertilized post-meiosis I diploid oocytes obtained from each donor to develop multiple blastocyst-like masses, each of which contains an inner cell mass (ICM) that is homozygous for a particular MHC haplotype, wherein the non-fertilized post-meiosis I diploid oocytes are subject to sham intracytoplasmic sperm injection (ICSI) followed by incubation with calcium ionophore; (d) isolating HES cells from the ICM obtained from each donor; (e) culturing the isolated HES cells to obtain HES cell lines; (f) determining the MHC haplotype of each HES cell line; and (g) cataloging the MHC haplotype of each HES cell line obtained in (e).
Claim 2
Independent Claim
149 words
A method for producing a mammalian homozygous embryonic stem (HES) cell depository, wherein each HES cell produces a cell line, which has alleles that are homozygous for a Major Histocompatibility Complex (MHC) haplotype, wherein the method comprises: (a) selecting donors; (b) determining the MHC haplotype of each donor; (c) mitotically activating non-fertilized post-meiosis I diploid oocytes obtained from each donor to develop multiple blastocyst-like masses, each of which contains an inner cell mass (ICM) that is homozygous for a particular MHC haplotype, wherein the non-fertilized post-meiosis I diploid oocytes are subject to sham intracytoplasmic sperm injection (ICSI) followed by incubation with calcium ionophore; (d) isolating HES cells from the ICM obtained from each donor; (e) culturing the isolated HES cells to obtain HES cell lines; (f) determining the MHC haplotype of each HES cell line; and (g) cataloging the MHC haplotype of each HES cell line obtained in (e).
Claim 3
Dependent Claim
11 words
The method of claim 1 , wherein the donors are human.
Claim 4
Dependent Claim
11 words
The method of claim 1 , wherein the donors are non-human.