Transdifferentiation Potential of Bone Marrow Stem Cells

骨髓干细胞的转分化潜力

• 批准号: 6717692
• 负责人: Makio Ogawa
• 金额: $ 31.5万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6717692
• 关键词: bone marrow; cell differentiation; cell population study; clone cells; confocal scanning microscopy; flow cytometry; fluorescence microscopy; green fluorescent proteins; hematopoietic stem cells; immunomagnetic separation; interference microscopy; laboratory mouse; regeneration; stem cell transplantation; tissue /cell culture

DESCRIPTION (provided by applicant): Recent studies demonstrated that stem cells in a number of organs/tissues have the potential to differentiate into tissues other than those specified in their origins. The most striking evidence for this "transdifferentiation" has been between bone marrow and muscular, hepatic, and CNS systems. Since all of the transplantation studies have been conducted using populations of cells, it is possible that the apparent bipotentiality of the stem cells is from engraftment by two separate populations of monopotential stem cells. Our proposal seeks definitive information on the tissue/organ engrafting capabilities of the hematopoietic and stromal stem cells by using single stem cell transplantation. We will use transgenic CD45.2 C57BL/6 (B6) mice that ubiquitously express enhanced green fluorescent protein (EGFP) as the source for donor stem cells and congenic CD45.1 B6 mice as recipients. The contribution of bone marrow EGFP-labeled donor cells to various organs/tissues of the recipient mice will be assessed using laser scanning confocal microscopy (LSCM) and conventional epifluorescence microscopy in combination with differential interference contrast (DIC) microscopy. Two Specific Aims are proposed: (1) To define the potentials of individual hematopoietic stem cells by single stem cell transplantation to lethally irradiated recipient mice. (2) To define the potentials of stromal stem cells. We have developed a highly efficient technique to create mice with clonal hematopoietic engraftment by combining FACS cell sorting and short-term suspension culture of bone marrow cells. Collaboration with the laboratory of Dr. Christopher J. Drake of Department of Cell Biology and Anatomy, who has considerable expertise in the use of LSCM and DIC and epifluorescence microscopy, has already resulted in exciting preliminary information about the potentiality of a single EGFP-labeled stem cell.

描述（由申请人提供）：最近的研究表明，许多器官/组织中的干细胞有可能分化成除其起源中指定的组织之外的组织。这种“转分化”最引人注目的证据是在骨髓和肌肉、肝脏和中枢神经系统之间。由于所有移植研究都是使用细胞群进行的，因此干细胞的表观双潜能可能来自两个独立的单能干细胞群的植入。我们的提案寻求通过使用单干细胞移植获得造血干细胞和基质干细胞的组织/器官移植能力的明确信息。我们将使用普遍表达增强型绿色荧光蛋白 (EGFP) 的转基因 CD45.2 C57BL/6 (B6) 小鼠作为供体干细胞的来源，并使用同源 CD45.1 B6 小鼠作为受体。将使用激光扫描共聚焦显微镜（LSCM）和传统落射荧光显微镜结合微分干涉对比（DIC）显微镜来评估骨髓EGFP标记的供体细胞对受体小鼠的各种器官/组织的贡献。提出了两个具体目标：（1）通过将单个干细胞移植到经致死照射的受体小鼠中来确定单个造血干细胞的潜力。 (2) 明确基质干细胞的潜力。我们开发了一种高效的技术，通过结合 FACS 细胞分选和骨髓细胞短期悬浮培养来创建具有克隆造血移植的小鼠。与细胞生物学和解剖学系的 Christopher J. Drake 博士的实验室合作，他在 LSCM 和 DIC 以及落射荧光显微镜的使用方面拥有丰富的专业知识，已经获得了关于单个 EGFP 标记的潜力的令人兴奋的初步信息干细胞。