异种肝前体细胞移植治疗急性肝损伤大鼠

背景：成体肝前体细胞可在受体肝脏内定植并分化为肝细胞。不过，异种肝前体细胞移植能否促进急性肝损伤的恢复，脾脏微环境能否促进移植物的存活和向肝细胞分化，尚没有研究。目的：评价异种肝前体细胞移植治疗急性肝损伤的作用；监测移植肝前体细胞在大鼠脾脏实质内的定植及向肝细胞的分化。方法：体外培养雄性小鼠来源的肝前体细胞系肝上皮样前体细胞。通过CCl4腹腔注射联合2/3肝切除构建急性肝损伤大鼠模型，进行肝上皮样前体细胞脾脏移植。在肝切除后1，5，14和21 d，苏木精-伊红染色观察肝脏病理改变，全自动生化分析仪监测血清转氨酶变化，PCR反应检测脾脏组织Y染色体特异性序列Sry，脾脏CK-19和Alb免疫组织化学追踪移植肝上皮样前体细胞的植入和肝细胞分化。结果与结论：肝上皮样前体细胞可在体外长期培养，保持增殖能力和双向分化潜能。肝上皮样前体细胞脾脏移植后，肝损伤大鼠肝细胞肿胀明显减轻，丙氨酸转氨酶和天门冬氨酸转氨酶下降更明显。移植后1，5，14和21 d，脾脏DNA中均能检测到Sry序列。在整个实验期间CK-19阳性细胞在大鼠脾脏实质内始终存在。Alb阳性细胞在移植后5 d在脾脏实质中出现，随后阳性细胞数逐渐增多。实验表明，移植肝前体细胞能在大鼠脾脏实质中植入，并分化为肝细胞，能有效促进CCl4腹腔注射联合2/3肝切除诱导的大鼠急性肝损伤的修复过程。

Intrasplenic delivery of xenogeneic hepatic progenitor cells ameliorates acute liver injury in rats

BACKGROUND： Adult hepatic progenitor cells transplanted into the liver can engraft and differentiate into hepatocytes. However, little evidence was available concerning whether heterogeneic hepatic progenitor cells can promote the restoration of acute liver damage and whether the microenvironment of spleen can promote the graft survival and differentiation into hepatocytes. OBJECTIVE： To evaluate the effect of hepatic progenitor cell transplantation on acute liver injury and monitor the engraftment and hepatocytic differentiation of transplanted hepatic progenitor cells in the splenic parenchyma.METHODS： Adult mouse derived progenitor cell line liver epithelial progenitor cells were cultured in vitro for long term. The acute liver injury model was established by intraperitoneal injection of CCl4 in combination with 2/3 partial hepatectomy, for transplantation of liver epithelial progenitor cells. The liver pathological changes were observed by hematoxylin-eosin staining at 1, 5, 14 and 21 days after hepatectomy. Serum transaminase levels were monitored using automatic biochemistry analyzer. The Sry sequence in the spleens was tested by PCR.Immunohistochemistry staining against CK-19 and Alb was performed to analyze the engraftment and hepatocytic ifferentiation of transplanted hepatic progenitor cells in the splenic parenchyma. BACKGROUND： Adult hepatic progenitor cells transplanted into the liver can engraft and differentiate into hepatocytes. However, little evidence was available concerning whether heterogeneic hepatic progenitor cells can promote the restoration of acute liver damage and whether the microenvironment of spleen can promote the graft survival and differentiation into hepatocytes. OBJECTIVE： To evaluate the effect of hepatic progenitor cell transplantation on acute liver injury and monitor the engraftment and hepatocytic differentiation of transplanted hepatic progenitor cells in the splenic parenchyma.METHODS： Adult mouse derived progenitor cell line liver epithelial progenitor cells were cultured in vitro for long term. The acute liver injury model was established by intraperitoneal injection of CCl4 in combination with 2/3 partial hepatectomy, for transplantation of liver epithelial progenitor cells. The liver pathological changes were observed by hematoxylin-eosin staining at 1, 5, 14 and 21 days after hepatectomy. Serum transaminase levels were monitored using automatic biochemistry analyzer. The Sry sequence in the spleens was tested by PCR.Immunohistochemistry staining against CK-19 and Alb was performed to analyze the engraftment and hepatocytic differentiation of transplanted hepatic progenitor cells in the splenic parenchyma. RESULTS AND CONCLUSION： Liver epithelial progenitor cells maintained normal proliferation and bipotential differentiation capacity after long-term culture in vitro. Compared with model group, hepatocyte edema was alleviated significantly, and serum alanine aminotransferase and aspartic transaminase levels were decreased remarkably after cell transplantation group. The Sry sequence was tested in spleen DNAs at 1, 5, 14 and 21 days post-transplantation. CK-19 positive cells existed in the splenic parenchyma throughout the study period. Alb-positive cells emerged at 5 days post-transplantation and the number of Alb-positive cells was gradually increased over time. Results show that hepatic progenitor cells engraft and differentiate into hepatocytes in rat splenic parenchyma successfully, and the transplantation can promote the recovery process from CCl4/partial hepatectomy induced acute liver injury.