Department of Electron Microscopy, Theodor Bilharz Research Institute, Giza, Egypt.
APMIS. 2010 Apr;118(4):277-87. doi: 10.1111/j.1600-0463.2010.02585.x.
The efficiency of differentiation of bone marrow cells (BMCs) into hepatocytes in vivo and its importance in physiopathological processes is still debated. Murine schistosomiasis was used as a liver injury model and unfractionated male mice BMCs were transplanted through intrahepatic injection into non-irradiated Schistosoma mansoni-infected female mice on their 16th week post-infection. Two weeks after bone marrow transplantation, mice were sacrificed on a weekly basis until 10 weeks. Tracing of male donor-derived cells in female recipient mice livers was carried out by the detection of Y chromosome expression by fluorescent in situ hybridization (FISH) and also of chromodomain Y-linked (CDYL) protein by indirect immunofluorescence (IF). Their transformation into hepatocytes was studied by double labelling indirect IF using antibodies directed against CDYL and mouse albumin. Histopathological and electron microscopic examinations revealed the presence of small hepatocyte-like cells in the periportal tracts and in between the hepatocytes facing the sinusoids. Donor-derived cells showing Y chromosome by FISH and expressing CDYL protein by IF were recovered in the infected transplanted livers. The initial number of these cells increased with increased post-transplantation time. Cells were mainly localized in the periphery of schistosoma granuloma. Few donor-derived cells appeared within the hepatic parenchymal tissue and showed positivity for albumin secretion by double labelling with IF. We suggest that transplanted bone marrow stem cells can repopulate the Schistosoma-infected liver of immunocompetent mice. Their differentiation is a complex event controlled by many factors and needs to be further characterized extensively. The extent and type of liver injury and the number of transplanted cells are important variables in the process of stem cell engraftment and differentiation into functioning hepatic cells that still need to be defined.
骨髓细胞(BMC)在体内分化为肝细胞的效率及其在生理病理过程中的重要性仍存在争议。利用小鼠日本血吸虫病作为肝损伤模型,将未分离的雄性小鼠 BMC 通过肝内注射移植到感染日本血吸虫的非照射雌性小鼠第 16 周后。骨髓移植后 2 周,每周处死小鼠,直至第 10 周。通过荧光原位杂交(FISH)检测 Y 染色体表达以及间接免疫荧光(IF)检测色氨酸结构域 Y 连锁(CDYL)蛋白,追踪雌性受者小鼠肝脏中雄性供体细胞。通过使用针对 CDYL 和小鼠白蛋白的双标记间接 IF 研究其向肝细胞的转化。组织病理学和电子显微镜检查显示,在门脉周围和面对窦状隙的肝细胞之间存在小的肝细胞样细胞。通过 FISH 显示 Y 染色体和 IF 表达 CDYL 蛋白的供体细胞在感染的移植肝脏中被回收。这些细胞的初始数量随着移植后时间的增加而增加。细胞主要定位于血吸虫肉芽肿的外围。少数供体细胞出现在肝实质组织内,并通过 IF 双标记显示白蛋白分泌阳性。我们认为,移植的骨髓干细胞可以重新填充免疫功能正常的小鼠感染的肝脏。它们的分化是一个受多种因素控制的复杂事件,需要进一步广泛地进行特征描述。肝损伤的程度和类型以及移植细胞的数量是干细胞植入和分化为有功能的肝细胞过程中的重要变量,仍需要进一步定义。