Biochemical and molecular characterization of hepatocyte-like cells derived from human bone marrow mesenchymal stem cells on a novel three-dimensional biocompatible nanofibrous scaffold
Biochemical and molecular characterization of hepatocyte-like cells derived from human bone marrow mesenchymal stem cells on a novel three-dimensional biocompatible nanofibrous scaffold
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DOI：
10.1111/j.1440-1746.2008.05530.x
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发表时间：
2009-02-01
影响因子：
4.1
通讯作者：
Jazayery, Maryam
中科院分区：
文献类型：
--
作者：
Kazemnejad, Somaieh;Allameh, Abdolamir;Jazayery, Maryam
纳米纤维在干细胞组织工程中的应用引起了人们极大的兴趣。间充质干细胞在纳米纤维结构中向肝系的转分化尚未见报道。本研究采用三维纳米纤维支架将人骨髓间充质干细胞（hBMSCs）分化为肝细胞。采用静电纺丝技术制备了由聚己内酯、胶原蛋白和聚醚砜组成的支架。对分离的hBMSCs进行表征后，通过扫描电镜（SEM）和MTT实验评估细胞在支架上的性能。细胞学、分子和生化指标的测定证实了造血干细胞向肝细胞分化的潜力。分离的细胞具有间充质干细胞的基本特性。扫描电镜（SEM）和MTT分析表明，细胞在纳米纤维上粘附、渗透和增殖。在纳米纤维上培养的细胞分化为肝细胞样细胞，并在mRNA水平上表达肝细胞特异性标志物，如白蛋白、甲胎蛋白、细胞角蛋白-18、细胞角蛋白-19和细胞色素P450 3A4。与二维培养系统（28 +/- 6%）相比，在支架上培养的白蛋白阳性细胞数量相当多（47 +/- 4%），这表明支架的支撑作用。细胞在支架上的肝细胞中产生白蛋白、尿素、转铁蛋白、血清谷丙转氨酶和血清草酰乙酸转氨酶的效率进一步证明了细胞的功能。本研究提供的数据表明，工程纳米纤维支架是一种导电基质，支持并促进间充质干细胞发育为功能性肝细胞样细胞。
There is significant interest in using nanofibers in tissue engineering from stem cells. The transdifferentiation of mesenchymal stem cells into the hepatic lineage in a nanofibrous structure has not been reported. In this study, a three dimensional nanofibrous scaffold is introduced for differentiation of human bone marrow derived mesenchymal stem cells (hBMSCs) into hepatocytes.A scaffold composed of Poly (epsilon-caprolactone), collagen and polyethersulfone was fabricated by the electrospinning technique. After characterization of isolated hBMSCs, the performance of the cells on the scaffold was evaluated by Scanning Electron Microscopy (SEM) and MTT assay. Cytological, molecular and biochemical markers were measured to confirm differentiation potential of hBMSCs into hepatocytes.The isolated cells possessed the basic properties of mesenchymal stem cells (MSCs). Based on scanning electron microscope (SEM) analysis and MTT assay, it was shown that the cells adhere, penetrate and proliferate on the nanofibers. Cultured cells on the nanofibers differentiated into hepatocyte-like cells and expressed hepatocyte specific markers such as albumin, alpha-fetoprotein, cytokeratin-18, cytokeratin-19 and cytochrome P450 3A4 at mRNA levels. Appearance of a considerable number of albumin-positive cells cultivated on the scaffold (47 +/- 4%) as compared to the two-dimensional culture system (28 +/- 6%) indicates the supporting role of the scaffold. The efficiency of the cells to produce albumin, urea, transferrin, serum glutamic pyruvic transaminase and serum oxaloacetate aminotransferase in hepatocytes on the scaffold further attest to the functionality of the cells.The data presented in this study show that the engineered nanofibrous scaffold is a conductive matrix which supports and enhances MSC development into functional hepatocyte-like cells.