Ren Hong-ying, Zhao Qin-jun, Xing Wen, Yang Shao-guang, Lu Shi-hong, Ren Qian, Zhang Lei, Han Zhong-chao
State Key Laboratory of Experimental Hematology, Institute of Hematology, CAMS and PUMC, Tianjin 300020, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2010 Apr;32(2):190-4. doi: 10.3881/j.issn.1000-503X.2010.02.013.
To investigate the biological function of hepatocyte-like cells derived from mesenchymal stem cells that isolated from human umbilical cord UC-MSCs in vitro, and to detect the changes in the immunogenicity of the differentiated hepatocyte-like cells (DHC).
Transdifferentiation of UC-MSCs into hepatic lineage in vitro was induced in modified two-step induction medium. The expressions of hepatic specific markers were detected by RT-PCR analysis and immunofluorescence staining at different time points after induction. The levels of albumin and urea in the supernatants of cultures were measured by enzyme-linked immunosorbent assay. Furthermore, the immunosuppressive property of DHC was detected by one-way mixed lymphocyte culture.
The mRNA and proteins of alpha fetoprotein (AFP), albumin (ALB),and cytokeratin-19 (CK-19) were expressed in naive UC-MSCs at low levels. DHC highly expressed hepatic markers AFP, ALB, CK-19, and tryptophan 2, 3-dioxygenase 14 and 28 days after hepatic differentiation and were accompanied by an increased production of ALB and urea in supernatant in a time-dependent manner. DHC did not express human leukocyte antigen DR antigen and significantly decreased the lymphocyte proliferation.
UC-MSCs are able to differentiate into functional hepatocyte-like cells in vitro, while the immunogenicity of DHC remains low.
研究从人脐带中分离的间充质干细胞(UC-MSCs)来源的肝细胞样细胞在体外的生物学功能,并检测分化的肝细胞样细胞(DHC)免疫原性的变化。
在改良的两步诱导培养基中诱导UC-MSCs体外向肝系分化。诱导后不同时间点通过逆转录聚合酶链反应(RT-PCR)分析和免疫荧光染色检测肝特异性标志物的表达。采用酶联免疫吸附测定法测量培养上清液中白蛋白和尿素的水平。此外,通过单向混合淋巴细胞培养检测DHC的免疫抑制特性。
甲胎蛋白(AFP)、白蛋白(ALB)和细胞角蛋白19(CK-19)的信使核糖核酸(mRNA)和蛋白质在未分化的UC-MSCs中低水平表达。肝分化后14天和28天,DHC高表达肝标志物AFP、ALB、CK-19和色氨酸2,3-双加氧酶,且上清液中ALB和尿素的产生呈时间依赖性增加。DHC不表达人白细胞抗原DR抗原,并显著降低淋巴细胞增殖。
UC-MSCs能够在体外分化为功能性肝细胞样细胞,而DHC的免疫原性仍然较低。