Khodabandeh Zahra, Vojdani Zahra, Talaei-Khozani Tahereh, Jaberipour Mansoureh, Hosseini Ahmad, Bahmanpour Soghra
Laboratory for Stem Cell Research, Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran ; Transgenic Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Laboratory for Stem Cell Research, Department of Anatomy, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Iran J Med Sci. 2016 Jan;41(1):28-36.
Human Wharton's jelly mesenchymal stem cells (HWJMSCs) express liver-specific markers such as albumin, alpha-fetoprotein, cytokeratin-19, cytokeratin-18, and glucose-6-phosphatase. Therefore, they can be considered as a good source for cell replacement therapy for liver diseases. This study aimed to evaluate the effects of various culture systems on the hepatocyte-specific gene expression pattern of naïve HWJMSCs.
HWJMSCs were characterized as MSCs by detecting the surface CD markers and capability to differentiate toward osteoblast and adipocyte. HWJMSCs were cultured in 2D collagen films and 3D collagen scaffolds for 21 days and were compared to control cultures. Real time RT-PCR was used to evaluate the expression of liver-specific genes.
The HWJMSCs which were grown on non-coated culture plates expressed cytokeratin-18 and -19, alpha-fetoprotein, albumin, glucose-6-phosphatase, and claudin. The expression of the hepatic nuclear factor 4 (HNF4) was very low. The cells showed a significant increase in caludin expression when they cultured in 3D collagen scaffolds compared to the conventional monolayer culture and 2D collagen scaffold.
Various culture systems did not influence on hepatocyte specific marker expression by HWJMSCs, except for claudin. The expression of claudin showed that 3D collagen scaffold provided the extracellular matrix for induction of the cells to interconnect with each other.
人脐带华通氏胶间充质干细胞（HWJMSCs）表达肝脏特异性标志物，如白蛋白、甲胎蛋白、细胞角蛋白-19、细胞角蛋白-18和葡萄糖-6-磷酸酶。因此，它们可被视为肝脏疾病细胞替代治疗的良好来源。本研究旨在评估各种培养系统对未处理的HWJMSCs肝细胞特异性基因表达模式的影响。
通过检测表面CD标志物以及向成骨细胞和脂肪细胞分化的能力，将HWJMSCs鉴定为间充质干细胞。将HWJMSCs在二维胶原膜和三维胶原支架中培养21天，并与对照培养物进行比较。采用实时逆转录聚合酶链反应（RT-PCR）评估肝脏特异性基因的表达。
在未包被的培养板上生长的HWJMSCs表达细胞角蛋白-18和-19、甲胎蛋白、白蛋白、葡萄糖-6-磷酸酶和紧密连接蛋白。肝细胞核因子4（HNF4）的表达非常低。与传统单层培养和二维胶原支架相比，当细胞在三维胶原支架中培养时，紧密连接蛋白的表达显著增加。
除紧密连接蛋白外，各种培养系统均未影响HWJMSCs的肝细胞特异性标志物表达。紧密连接蛋白的表达表明三维胶原支架为诱导细胞相互连接提供了细胞外基质。
