Department of Medicine, Division of Hematology and Oncology, Rhode Island Hospital and Brown University Medical School, Providence, Rhode Island 02903, USA.
J Histochem Cytochem. 2010 Jan;58(1):61-72. doi: 10.1369/jhc.2009.954560. Epub 2009 Sep 28.
In the present report, we have compared the phenotype and growth of small hepatocyte progenitors (SHPs) induced by retrorsine/partial hepatectomy (R/PH) and small hepatocytes (SHs) isolated from normal adult liver. SHs were isolated by a combination of differential centrifugation and Percoll isodensity fractionation from a liver cell suspension prepared by collagenase perfusion of a dipeptidyl peptidase IV (DPPIV)-positive Fischer F344 rat liver. Following further purification by flow cytometry, the SH-R3 fraction was transplanted via the portal vein into R/PH-treated, DPPIV-negative Fischer F344 rats. Frozen sections from tissue harvested at 5, 7, and 21 days after transplantation were analyzed by indirect immunofluorescence to compare the phenotypic characteristics of colonies formed by exogenous SH-R3s and endogenous SHPs. Colonies of transplanted SHs and endogenous SHPs displayed similar histologies and phenotypes but were distinguished from surrounding hepatocytes by their elevated expression of transferrin receptor. SH-R3 colonies were frequently located within clusters of gamma-glutamyl transpeptidase-positive host hepatocytes. Although significantly smaller at 5 and 7 days after PH, by day 21, SH-R3 colonies were similar in size to those formed by SHPs. The present results suggest that endogenous SHPs are derived, at least in part, from SHPs.
在本报告中，我们比较了反式丝氨酸/部分肝切除术（R/PH）诱导的小肝细胞祖细胞（SHPs）和从正常成年肝脏中分离的小肝细胞（SHs）的表型和生长情况。SHs 是通过胶原酶灌注二肽基肽酶 IV（DPPIV）阳性 Fischer F344 大鼠肝脏制备的肝细胞悬浮液，通过差速离心和 Percoll 等密度分级分离来分离。通过流式细胞术进一步纯化后，将 SH-R3 级分通过门静脉移植到 R/PH 处理的、DPPIV 阴性的 Fischer F344 大鼠中。在移植后 5、7 和 21 天采集的组织冷冻切片通过间接免疫荧光分析，比较由外源性 SH-R3 和内源性 SHPs 形成的集落的表型特征。移植的 SHs 和内源性 SHPs 集落显示出相似的组织学和表型，但通过转铁蛋白受体的高表达与周围肝细胞区分开来。SH-R3 集落经常位于γ-谷氨酰转肽酶阳性宿主肝细胞簇内。尽管在 PH 后 5 和 7 天明显较小，但到第 21 天，SH-R3 集落的大小与 SHPs 形成的集落相似。目前的结果表明，内源性 SHPs 至少部分来源于 SHPs。
