ORIGIN AND ISOLATION OF LIVER PROGENITOR CELLS

描述(由申请人提供)：最近的研究已经确认在成人器官中存在干细胞。 然而，肝干细胞尚未被鉴定和分离。在……下面 在某些情况下，当肝细胞增殖受损时，肝祖细胞 细胞，也被称为椭圆形细胞，在肝脏中出现、增殖和开始 沿肝细胞或胆管谱系分化。最近的研究还表明， 表明骨髓细胞或纯化的造血干细胞能够 在肝脏中分化为肝细胞和胆汁上皮细胞。我们的 工作假说是肝脏祖细胞/卵圆细胞是两者的后代 肝脏和造血干细胞。因为伟大的生物医学 这些细胞的重要性，我们的长期目标是分离这些细胞 细胞，在适当的条件下体外扩增，并用于 动物模型中病变肝脏的再生和治疗。要实现 为了实现这一目标，确定它们的原产地是至关重要的，无论是 它们来自肝脏、骨髓或这两个器官。至 确定祖细胞/卵圆细胞是否从基因上来源于骨髓 标记的骨髓细胞将被移植到致命性照射的 受体及其在D-氨基半乳糖中的形态和分布 逆皮质醇/部分肝切除损伤模型将被监测。使用已知 椭圆形细胞标记物，我们将能够确定一些 移植细胞的行为类似于肝脏祖细胞，并开始 沿肝细胞/胆管谱系分化。如果肝脏祖细胞 都是骨髓细胞的后代，鉴定将是极其重要的 通过设计适当的细胞标志物的组合来表达它们的前体 在造血祖细胞亚群和椭圆形细胞中唯一表达 细胞，并使用这组标记将它们从骨髓中分离出来。跟随 同样的策略，我们将为 肝脏中祖细胞的鉴定。记号笔的面板将 代表在造血祖细胞中表达的细胞表面标志， 肝母细胞、胆管上皮细胞和肝细胞。这两者的结合 将设计出仅在肝脏祖细胞/卵圆形细胞中表达的标记。 这组标志物随后将用于分离肝脏祖细胞 取自肝脏。

DESCRIPTION (Provided by Applicant): Recent research has recognized the existence of stem cells in adult organs. However, liver stem cells have not yet been identified and isolated. Under certain conditions, when hepatocyte proliferation is impaired, liver progenitor cells, also termed oval cells, emerge in the liver, proliferate and commence differentiationalong hepatocyticor bile duct lineages. Recent studies have also shown that bone marrow cells or purified hematopoietic stem cells are capable of differentiating in the liver into hepatocytes and bile epithelial cells. Our working hypothesis is that liver progenitor/oval cells are progeny of both liver and hematopoietic stem cells. Because of the great biomedical significance of these cells, our long-term objectives are to isolate these cells, expand them in vitro under appropriate conditions and use them for repopulation and treatment of the diseased liver in animal models. To achieve this goal, it is of great importance to identify their site of origin, whether they originate from the liver, from bone marrow or both of these organs. To determine whether progenitor/oval cells originate from bone marrow, genetically marked bone marrow cells will be transplanted into lethally irradiated recipients and their appearance and distribution in D-galactosamine and retrorsine/partial hepatectomy injury models will be monitored. Using known oval cell markers, we will be able to determine whether some of the transplanted cells behave like liver progenitor cells and commence differentiation along hepatocytic/bile duct lineages. If liver progenitor cells are progeny of bone marrow cells, it will be extremely important to identify their precursors by designing a combination of appropriate cell markers uniquely expressed in a subset of hematopoietic progenitor cells and the oval cells and use this panel of markers to isolate them from bone marrow. Following the same strategy, we will design a combination of markers for the identification of progenitor cells in the liver. The panel of markers will represent cell surface markers expressed in hematopoietic progenitor cells, hepatoblasts, biliary epithelial cells and hepatocytes. A combination of these markers will be designed that is expressed only in liver progenitor/oval cells. This panel of markers will be then used for isolation of liver progenitor cells from the liver.