Proliferation and differentiation of hepatic stem cell from bone marrow cells and possibility of bone marrow transplantation in genetic hepatic disease

全肝或部分肝移植是治疗人类终末期肝病的唯一有效方法，但其应用受到供体器官数量不足和成本高的限制。异体肝细胞移植作为肝移植的一种替代方法已被研究。但它的使用也受到捐助者数量有限的限制。近年来的许多研究表明，骨髓细胞在体外和体内可分化为多种细胞系，如脂肪细胞、心肌细胞、皮肤上皮细胞、神经元细胞和肝细胞。此外，Lagasse等人发现骨髓细胞移植到富马酰乙酸水解酶（FAH）缺陷小鼠（人类致死性先天性酪氨酸血症型动物模型）中，可以挽救小鼠并恢复其肝功能，Li等人在颈动脉内给予骨髓基质细胞[6]，可以改善大鼠中风模型的神经功能恢复。越来越多的研究表明，骨髓细胞是细胞移植治疗的理想资源。[2003]我们研究了异体骨髓细胞移植治疗长酶无球蛋白血症大鼠（NAR）先天性白蛋白缺失的可行性。7周龄雄性NARs作为受体，6 ~ 8周龄雄性SD大鼠作为同种异体移植供体。NARs分为三组：骨髓移植（BMT）组（n=10），骨髓细胞输注于肝脏；肝细胞移植（HCT）组（n=8），肝细胞移植入肝脏；对照组（n=8）门静脉注射PBS。测定血清白蛋白水平作为移植细胞功能的指标，并利用免疫组织化学和免疫荧光技术评估移植细胞在受体肝脏中的表型特征。细胞移植后8周，BMT组和HCT组血清白蛋白水平显著高于对照组。来源于骨髓细胞的肝细胞样细胞在NARs的肝脏中表达白蛋白。由此可见，骨髓细胞在体内可分化为肝细胞样细胞。结果表明，骨髓细胞移植是治疗先天性白蛋白血症模型大鼠的有效方法，提示同种异体骨髓细胞移植可作为一种有效的治疗遗传性代谢疾病的方法。<2004>最近的研究表明，链脲佐菌素诱导的糖尿病后骨髓细胞移植可以支持胰腺β细胞质量的恢复和高血糖的部分逆转。为了解决这个问题，我们研究了c-Met/肝细胞生长因子（HGP）信号通路是否参与骨髓移植（BMT）后β细胞损伤的恢复。在该模型中，供体来源的骨髓细胞在受体脾、肝、肺和胰腺以及宿主肝细胞中HGF免疫反应性呈阳性。事实上，血浆HGF水平维持在较高水平。BMT组胰腺导管细胞中c-Met的表达频率、增殖活性和分化反应均高于pbs处理组，导致内源性胰岛素生成细胞数量增加。BMT诱导c-Met/HGF信号通路促进糖尿病大鼠胰腺再生。少

Whole liver or partial liver transplantation is the only effective method of treating end-stage liver disease in humans, but its application is limited by an insufficient number of donor organs and the high cost. Allogeneic hepatocyte transplantation has been studied as an alternative method of liver transplantation. But its use also has been limited by the limited number of donors.Many studies in the past several years have shown that bone marrow cells can differentiate into various cell lineage in vitro and in vivo such as lipocyte, cardiac myoblast, skin epithelium, neuron, and hepatocytes lineage. Moreover, Lagasse, et. al. found that bone marrow cell transplantation into fumarylacetoacetate hydrolase (FAH)-deficient mice, an animal model of human fatal congenital tyrosinemia type rescued the mice and restored their liver function, and Li, et. Al. showed improvement of neurologic recovery in a rat stroke model with intracarotid administration of bone marrow stromal cell[6]. These s … More tudies demonstrated that bone marrow cells are an ideal resource for cell transplantation therapy.<2003>We investigated the feasibility of correcting the congenital absence of albumin in Nagase analbuminemic rats (NAR) by allogeneic bone marrow cell transplantation. Seven week-old male NARs were used as recipients, and 6- to 8- week-old male Sprague-Dawley (SD) rats were used as allograft donors. NARs were divided into three groups: a bone marrow transplantation (BMT) group (n=10) in which bone marrow cells were infused into the liver; a hepatocyte transplantation (HCT) group (n=8) in which hepatocytes were transplanted into the liver; and a control group (n=8) in which PBS was injected into the portal vein. Serum albumin levels were measured as an indicator of the function of the grafted cells, and the phenotypic characteristics of the engrafted cells in the recipient's liver were assessed with immunohistochemical and immunofluorescence techniques. At 8 weeks after cell transplantation the serum albumin levels of the BMT group and HCT group were significantly higher than in the control group. The hepatocyte-like cells derived from bone marrow cells expressed albumin in liver of the NARs. From this result, bone marrow cells can differentiate into hepatocyte-like cells in vivo. The results show that bone marrow cell transplantation is an effective treatment for congenital analbuminemia in a rat model and suggest that allogeneic bone marrow cell transplantation can be used as an efficient therapy for hereditary metabolic diseases.<2004>Recent studies have demonstrated that the transplantation of bone marrow cells following diabetes induced by streptozotocin can support the recovery of pancreatic beta-cell mass and a partial reversal of hyperglycemia. To address this issue, we examined whether the c-Met/hepatocyte growth factor (HGP) signaling pathway was involved in the recovery of beta-cell injury after bone marrow transplantation (BMT). In this model, donor-derived bone marrow cells were positive for HGF immunoreactivity in the recipient spleen, liver, lung, and pancreas as well as in the host hepatocytes. Indeed, plasma HGF levels were maintained at a high value. The frequency of c-Met expression and its proliferative activity and differentiative response in the pancreatic ductal cells in the BMT group were greater than those in the PBS-treated group, resulting in an elevated number of endogenous insulin-producing cells. The induction of the c-Met/HGF signaling pathway following BMT promotes pancreatic regeneration in diabetic rats. Less