DIFFERENTIATION OF BLOOD STEM CELLS INTO LIVER CELLS

描述：(改编自调查者的摘要)假设 骨髓中的祖细胞(骨髓源性肝祖细胞 细胞，BMLP)可以产生肝细胞，将在体内进行正式测试 在试管中。在具体目标1中，我们提出了一系列六个实验，其中 骨髓或肝脏来源的干细胞被移植到接受者体内。这些 实验将定义：1)激活供体中BMLP的条件。 2)可以提高接受者承兑能力的条件 培养BMLP。3)BMLPs与肝源性肝脏的关系 祖细胞。4)肝母细胞系产生 骨髓前体细胞。5)检测移植细胞的最佳标记。 供体小鼠将包括正常雄性、LacZ雄性和EGFP雄性小鼠。供体细胞 在接受辐射的女性接受者中将被检测到 Y染色体和B-半乳糖或绿色荧光蛋白。在特定目标2中，骨髓或血细胞 将在已知对肝母细胞有选择性的条件下进行培养，以 确定是否可以产生具有肝细胞表型的集落。在这两个地方 特异靶1和特异靶2，分离骨髓和血细胞 来自正常小鼠和部分肝切除或可卡因后的小鼠 诱导门静脉周围损伤、诱导肝细胞损伤的实验模型 肝干细胞增殖或门静脉周围肝干细胞增殖。这些 众所周知，手法可以诱导肝细胞和肝干细胞的生长因子 细胞，并可能鼓励肝脏对HSCs的测定。此外，尝试将 从骨髓或肝脏中分离出“前祖细胞” 可能具有多线作战的潜力。在体外克隆将被鉴定 形态和传代后分析细胞分化潜能 移植，表型表达，生长特性，对 分化刺激和超微结构。类似的体外研究将会是 在人的骨髓和血液上使用来自 骨髓移植实验室，否则就会被丢弃。如果这个 如果项目成功，它将克服可能使用的一个主要障碍 肝干细胞用于治疗肝功能衰竭或体外基因治疗。

DESCRIPTION: (Adapted from the investigator's abstract) The hypothesis that progenitor cells in the bone marrow (bone marrow derived liver progenitor cells, BMLP) can give rise to liver cells will be formally tested in vivo and in vitro. In specific Aim 1 we propose a series of six experiments in which bone marrow or liver derived stem cells are transplanted into recipients. These experiments will define: 1) The conditions which activate BMLPs in the donor. 2) The conditions which may increase the ability of the recipient to accept and to nurture BMLPs. 3) The relationship of BMLPs to liver derived liver progenitor cells. 4) The ability of an hepatoblast cell line to give rise to bone marrow precursors. 5) Optimal markers for detection of transplanted cells. Donor mice will include normal male, lacZ male and EGFP male mice. Donor cells in irradiated female recipients will be detected by the presence of Y-chromosome and B-gal or EGFP. In specific Aim 2, bone marrow or blood cells will be cultured, using conditions known to be selective for hepatoblasts, to determine if colonies with hepatocyte phenotype can be produced. In both Specific Aim 1 and specific Aim 2, bone marrow and blood cells will be isolated from both normal mice and mice following partial hepatectomy or after cocaine induced periportal injury, experimental models to induce hepatocyte proliferation or periportal liver stem cell proliferation, respectively. These manipulations are known to induce growth factors for hepatocytes and liver stem cells and may encourage liver determination of HSCs. In addition, attempts will be made to isolate "pre-progenitor" cells from the bone marrow or liver that may have multilinege potential. In vitro colonies will be identified morphologically and passaged for analysis of differentiation potential after transplantation, phenotypic expression, growth properties, response to differentiating stimuli, and ultrastructure. Similar in vitro studies will be done on human bone marrow and blood using reserve cell preparations from the bone marrow transplant laboratory, which would otherwise be discarded. If this project is successful, it would overcome a major hurdle in the possible use of hepatic stem cells for treatment of liver failure or ex vivo gene therapy.