Hematopoietic potential of histocompatible embryonic stem cell lines

组织相容性胚胎干细胞系的造血潜力

• 批准号: 8534806
• 负责人: Kitai Kim
• 金额: $ 23.27万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-25 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8534806
• 关键词: Affect; Blood; Bone Marrow; Cell Line; Cells; Childhood Leukemia; Chromosome Transfer; Chromosomes, Human, Pair 17; Chromosomes, Human, Pair 6; Development; Diploidy; ES Cell Line; Embryo; Epigenetic Process; Excision; Funding; Generations; Genetic Materials; Goals; Hand; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hereditary Disease; Histocompatibility Testing; Human; Hybrids; In Vitro; Instruction; Letters; Major Histocompatibility Complex; Mediating; Methods; Methylation; Modeling; Modification; Mus; Oocytes; Other Genetics; Patients; Phenotype; Pluripotent Stem Cells; Research; Research Subjects; Role; Schedule; Skin; Somatic Cell; Source; Staging; Stem cells; Supplementation; Techniques; Testing; Therapeutic; Tissues; Transplantation; Umbilical Cord Blood; cell type; design; embryonic stem cell; hematopoietic tissue transplantation; human tissue; in vivo; induced pluripotent stem cell; methylome; morphogens; novel; nuclear transfer; stem; tool; transcription factor

PROJECT SUMMARY (See instructions): The goal of this research is to create histocompatible patient specific hematopoietic stem cells, which in addition to their potential therapeutic value, will provide novel tools for in-depth study of reprogramming and differentiation. The Specific Aims of this proposal are: Specific Aims 1: Generation of histocompatible mmctES cells using microcell mediated chromosome transfer (mmct) in mouse. Specific Aim 2: Testing of hematopoietic potential in mmctES cells by in vitro and in vivo methods. The limited availability of human oocytes required for both nuclear transferred (nt) ES and parthenogenetic (p) ES cell techniques poses a significant obstacle to generating customized embryonic stem cells. Therefore, alternative methods to generate similar types of cells are needed. Generation of ES cells by oocyte free methods already has been demonstrated by fusion of somatic cells (sc) and embryonic stem (es) cells to reprogram the somatic cells. However, removal of the genetic material from the resultant tetraploid sc-es hybrid ES cells to make diploid ES cells has not been achieved. As an alternative approach, I propose microcell-mediated transfer to ES cells of mouse sc-chromosome 17 and human sc-chromosome 6, which contain the major histocompatibility complex (MHC). The resultant duplicated mouse es-chromosome 17 and human sc-chromosome 6 will then be removed to make diploid mmctES cells in mouse

项目总结(见说明)： 这项研究的目标是创造组织相容的患者特异性造血干细胞，这种干细胞除了具有潜在的治疗价值外，还将为深入研究重编程和分化提供新的工具。该方案的具体目标是：特定目标1：利用小鼠微细胞介导的染色体转移(MMCT)产生组织相容的MMCTES细胞。特定目的2：用体外和体内方法检测MMCTES细胞的造血潜能。核移植(NT)ES和孤雌生殖(P)ES细胞技术所需的人类卵母细胞的可获得性有限，这对生成定制的胚胎干细胞构成了重大障碍。因此，需要其他方法来生成类似类型的单元格。体细胞(Sc)和胚胎干细胞(ES)融合以重新编程体细胞，已经证明了用无卵母细胞的方法产生ES细胞。然而，从得到的四倍体SC-ES杂交ES细胞中去除遗传物质以形成二倍体ES细胞还没有实现。作为一种替代方法，我建议将含有主要组织相容性复合体(MHC)的小鼠sc-17号染色体和人sc-6号染色体的微细胞介导的转移到ES细胞。然后将复制的小鼠ES-17号染色体和人类SC-6号染色体移出，在小鼠体内产生二倍体的MmctES细胞。