Suzuki Kaori, Tanaka Minoru, Watanabe Natsumi, Saito Shigeru, Nonaka Hidenori, Miyajima Atsushi
Institute of Molecular and Cellular Biosciences, University of Tokyo, Tokyo, Japan.
Gastroenterology. 2008 Jul;135(1):270-281.e3. doi: 10.1053/j.gastro.2008.03.075. Epub 2008 Apr 4.
BACKGROUND & AIMS: Hepatic stellate cells (HSCs) and portal fibroblasts (PFs) are 2 distinct mesenchymal cells in adult liver. HSCs in sinusoids accumulate lipids and express p75 neurotrophin receptor (p75NTR). HSCs and PFs play pivotal roles in liver regeneration and fibrosis. However, the roles of mesenchymal cells in fetal liver remain poorly understood. In this study, we aimed to characterize mesenchymal cells in mouse fetal liver.
We prepared an anti-p75NTR monoclonal antibody applicable for flow cytometry and immunohistochemistry. p75NTR(+) cells isolated from fetal liver by flow cytometry were characterized by reverse-transcription polymerase chain reaction, immunohistochemistry, and cell cultivation. Lipid-containing cells were visualized by Oil-red O staining.
p75NTR(+) cells in fetal liver were clearly distinct from endothelial cells and showed characteristics of mesenchymal cells. At embryonic day (E) 10.5, p75NTR(+) cells were present at the periphery of the liver bud in close contact with endothelial cells, and spread over the liver at E11.5. With the formation of the liver architecture, they began to localize to 2 distinct areas, parenchymal and portal areas, and lipid-containing p75NTR(+) cells increased accordingly. p75NTR(+) cells around portal veins were adjacent to cholangiocytes and expressed Jagged1, a crucial factor for the commitment of hepatoblasts to cholangiocytes. By cultivation, p75NTR(+) cells showed features of adult HSCs with markedly increased expression of glial fibrillary acidic protein and alpha-smooth muscle actin.
p75NTR(+) mesenchymal cells in fetal liver include progenitors for HSCs and PFs, and the anti-p75NTR monoclonal antibody is useful for their isolation.
肝星状细胞（HSCs）和门周成纤维细胞（PFs）是成年肝脏中两种不同的间充质细胞。肝血窦中的肝星状细胞积累脂质并表达p75神经营养因子受体（p75NTR）。肝星状细胞和门周成纤维细胞在肝脏再生和纤维化中起关键作用。然而，间充质细胞在胎儿肝脏中的作用仍知之甚少。在本研究中，我们旨在对小鼠胎儿肝脏中的间充质细胞进行特征描述。
我们制备了一种适用于流式细胞术和免疫组织化学的抗p75NTR单克隆抗体。通过流式细胞术从胎儿肝脏中分离出的p75NTR(+)细胞，采用逆转录聚合酶链反应、免疫组织化学和细胞培养进行特征分析。含脂质的细胞通过油红O染色进行可视化。
胎儿肝脏中的p75NTR(+)细胞与内皮细胞明显不同，并表现出间充质细胞的特征。在胚胎第（E）10.5天，p75NTR(+)细胞出现在肝芽周边，与内皮细胞紧密接触，并在E11.5时扩散至整个肝脏。随着肝脏结构的形成，它们开始定位于两个不同区域，即实质区和门周区，含脂质的p75NTR(+)细胞也相应增加。门静脉周围的p75NTR(+)细胞与胆管细胞相邻，并表达Jagged1，这是肝母细胞向胆管细胞分化的关键因子。通过培养，p75NTR(+)细胞表现出成年肝星状细胞的特征，胶质纤维酸性蛋白和α-平滑肌肌动蛋白的表达明显增加。
胎儿肝脏中的p75NTR(+)间充质细胞包括肝星状细胞和门周成纤维细胞的祖细胞，抗p75NTR单克隆抗体有助于它们的分离。
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