STEM CELLS AND AGEING

干细胞与衰老

• 批准号: 7121494
• 负责人: STEWART SELL
• 金额: $ 24.36万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7121494
• 关键词: aging; bone marrow; bone marrow transplantation; cell migration; flow cytometry; green fluorescent proteins; hematopoietic stem cells; laboratory mouse; laser capture microdissection; p53 gene /protein; phenotype; sex chromosomes; stem cell transplantation

DESCRIPTION (provided by applicant): The hypothesis that bone marrow stem cells are a determining factor in ageing will be tested in a mouse model of premature ageing. First, the number and homing capacity of bone marrow stem cells from wild type p53+/+ and premature ageing p53+/m C57BL/6J mice will be assayed in vitro and in vivo. Then, both the ability of wild-type bone marrow stem cells to correct premature ageing in p53+/m mutant mice, and the ability of bone marrow stem cells from premature ageing p53+/m mutant mice to induce premature ageing, in wild-type recipients will be determined. In Specific Aim 1, the number of stem cells in wild-type p53+/+, p53+/- and p53+/m mice will be determined by flow cytometry and by in vitro colony formation, and homing determined by in vitro migration and marrow repopulation in vivo. In Specific Aim 2 bone marrow cells from GFP+ male C57BI6 p53+/+ mice will be transplanted to p53 mutant (p53+/m) and to wild-type (p53 +/+) female mice and the effect on the development of the aging phenotype determined. The contribution of donor cells to recipient tissues will be identified by the presence of GFP and the Y-chromosome. If ageing is due to a decreased ability of bone marrow stem cells to contribute to replacement of normal tissues in this model, transplantation of bone marrow cells from normal mice to p53 mutant mice should restore normal phenotype and normal life span. In Specific Aim 3 bone marrow cells from GFP+ p53+/m male mutant mice will be transplanted to irradiated normal wild-type female mice and to female mutant mice to determine if the aging phenotype can be transferred mutant bone marrow stem cells. Finally the possibility of fusion of donor bone marrow cells with tissue cells of recipient mice and the presence of fused cells in tumors will be determined. The results of these experiments should provide critical proof for one of the most important hypothetical properties of adult stem cells.

描述（由申请人提供）：将骨髓干细胞是衰老的决定因素的假设，将在早熟的小鼠模型中测试。首先，将在体外和体内分析野生型p53+/+的骨髓干细胞的数量和归巢能力和早熟p53+/m C57BL/6J小鼠的数量和归巢能力。然后，将确定野生型骨髓干细胞纠正p53+/m突变小鼠中过早衰老的能力，以及从过早衰老p53+/m突变小鼠诱导过早衰老的骨髓干细胞在野生型受体中诱导过早衰老的能力。在特定的目标1中，野生型p53+/+，p53 +/-和p53+/m小鼠的干细胞数量将由流式细胞仪和体外菌落形成确定，并通过体外迁移和骨髓在体内确定的归纳。在特定的目标中，来自GFP+雄性C57BI6 p53+/+小鼠的骨髓细胞将被移植到p53突变体（p53+/m）和野生型（p53+/+）雌性小鼠，以及对确定衰变表型的发育的影响。 GFP和Y染色体的存在将确定供体细胞对受体组织的贡献。如果衰老是由于骨髓干细胞在该模型中有助于替换正常组织的能力降低，则从正常小鼠到p53突变小鼠的骨髓细胞的移植应恢复正常的表型和正常寿命。在特定的目标中，来自GFP+ p53+/m雄性突变小鼠的3个骨髓细胞将被移植到辐照的正常野生型雌性小鼠和雌性突变小鼠中，以确定是否可以转移衰老表型突变体骨髓干细胞。最后，将确定将供体骨髓细胞与受体小鼠组织细胞融合的可能性以及肿瘤中融合细胞的存在。这些实验的结果应为成年干细胞最重要的假设特性之一提供关键证明。