Differentiation and migration of bone marrow mesenchymal stem cells transplanted through the spleen in rats with portal hypertension.
Differentiation and migration of bone marrow mesenchymal stem cells transplanted through the spleen in rats with portal hypertension.
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DOI：
10.1371/journal.pone.0083523
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Zheng S
中科院分区：
文献类型：
--
作者：
Sun S;Chen G;Xu M;Qiao Y;Zheng S
本研究旨在探讨骨髓间充质干细胞（BMSCs）在体外向肝细胞样细胞分化的能力，以及干细胞能否在门静脉高压症伴终末期肝病患者的肝脏内迁移和种植。 从大鼠中分离骨髓间充质干细胞，并用肝细胞生长因子（HGF）和成纤维细胞生长因子-4（FGF-4）扩增。免疫荧光法检测细胞中甲胎蛋白（AFP）、细胞角蛋白18（CK-18）和白蛋白（ALB）的表达。将大鼠随机分为实验组（胆总管结扎）和对照组。注射荧光标记细胞后，在荧光显微镜下观察细胞分布。使用Image-Pro Plus 6.0软件评价积分光密度（IOD）和细胞分布评分。处死大鼠前测定门静脉压力。 经HGF和FGF-4诱导后，第五代细胞的高尔基体、内质网、核糖体和线粒体均显著增加。免疫荧光分析显示，诱导后的细胞表达AFP、CK-18和ALB，CM-Dil染色阳性率达95.5%。实验组门静脉压力明显高于对照组（18.04± 2.35vs.9.75 ± 1.40cmH2O，P<0.01）。实验组移植细胞的IOD也明显高于对照组（11.30±2.09×105 vs.2.93 ±0.88×105，p<0.01）。实验组细胞分布评分低于对照组（1.99±0.36 vs.2.36 ±0.27，P<0.05）。 HGF和FGF-4的组合诱导BMSCs分化为肝细胞样细胞，其表达AFP、CK-18和ALB。此外，BMSCs的募集（移植到脾脏中后）在门静脉高压症大鼠中得到改善。
The goals of this paper were to evaluate the differentiation of bone marrow mesenchymal stem cells (BMSCs) into hepatocyte-like cells in vitro, and to determine whether stem cells can migrate and plant into the liver with portal hypertension accompanied by the end-stage of liver disease. BMSCs were isolated from rats and amplified with hepatocyte growth factor (HGF) and fibroblast growth factor-4 (FGF-4). The expression of alpha-fetoprotein (AFP), cytokeratin 18 (CK-18), and albumin (ALB) was detected by immunofluorescence in induced cells. Rats were randomly divided into experimental (with common bile duct ligation) and control groups. After injection of fluorescence labeled cells, cell distribution was observed under a fluorescence microscope. The integrated optical density (IOD) and cell distribution scores were evaluated using Image-Pro Plus 6.0 software. The portal pressure was measured before the rats were killed. After being induced with HGF and FGF-4, the Golgi apparatus, endoplasmic reticulum, ribosomes, and mitochondria all significantly increased in the fifth generation cells. Immunofluorescent analysis showed that the induced cells expressed AFP, CK-18, and ALB. BMSCs were stained by CM-Dil, and the labeling rate was as high as 95.5%. The portal pressure in experimental group was much higher than that of the control group (18.04±2.35 vs. 9.75±1.40cm H2O p<0.01). The IOD of transplanted cells in the experimental group was also significantly higher than that of the control group (11.30±2.09×105 vs. 2.93±0.88×105, p<0.01). In addition, the cell distribution score in the experimental group was lower than that of the control group (1.99±0.36 vs. 2.36±0.27, P<0.05). The combination of HGF and FGF-4 induces the differentiation of BMSCs into hepatocyte-like cells, which express AFP, CK-18, and ALB. In addition, the recruitment of BMSCs (after transplantation in the spleen) was improved in rats with portal hypertension.
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