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CD105 阳性猪子宫内膜基质间充质干细胞具有向心肌样细胞和胰岛素分泌β细胞样细胞分化的潜能。

CD105 Porcine Endometrial Stromal Mesenchymal Stem Cells Possess Differentiation Potential Toward Cardiomyocyte-Like Cells and Insulin-Producing β Cell-Like Cells In Vitro.

机构信息

1 Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, Jinju, Republic of Korea.

2 Animal Biotechnology Division, National Institute of Animal Science, RDA, Wanju-gun, Jeollabuk-do, Republic of Korea.

出版信息

Reprod Sci. 2019 May;26(5):669-682. doi: 10.1177/1933719118786461. Epub 2018 Jul 9.

Abstract

Porcine mesenchymal stem cells (MSCs) are similar to human MSCs, hence considered a valuable model for assessing potential for cell therapy. Porcine adipose-derived MSCs (AD-MSCs) and endometrial stromal MSCs (EMSCs) displayed fibroblast-like morphology and were positive for MSC markers CD73, CD90, and CD105 and negative for hematopoietic markers CD34 and CD45. The EMSCs had similar or slightly higher growth rate compared to AD-MSCs, and similar percentage of cells of both EMSCs and AD-MSCs were at G0/G1 and G2/M phases; however, EMSCs had significantly ( P < .05) higher percentage of cells at S phase of cell cycle than AD-MSCs. Transdifferentiation ability to cardiomyocyte-like cells was confirmed in differentiated cells by the expression of lineage-specific marker genes such as DES, ACTA2, cTnT, and ACTC1 by real-time quantitative polymerase chain reaction (RT-qPCR). Furthermore, cardiomyocyte-specific protein markers cTnT and ACTC1 were expressed in completely differentiated cells. Endodermal differentiation capacity of EMSCs to pancreatic β cell-like cells was evident with the changes in morphology and the expression of β-cell-specific marker genes such as PDX1, GLUT2, SST, NKX6.1, PAX4, and NGN3 as analyzed by RT-qPCR. The differentiated cells secreted insulin and C-peptide upon glucose challenge and also they expressed insulin, PDX1, PAX4, NGN3, and GLUT2 at protein level as assessed by immunostaining confirming the successful differentiation to β cell-like cells. Porcine EMSCs possess all the characteristics of MSCs and are suitable model for studying molecular mechanisms of cellular differentiation.

摘要

猪间充质干细胞(MSCs)与人 MSCs 相似,因此被认为是评估细胞治疗潜力的有价值模型。猪脂肪来源的 MSCs(AD-MSCs)和子宫内膜基质 MSCs(EMSCs)呈成纤维细胞样形态,并且 MSC 标志物 CD73、CD90 和 CD105 阳性,造血标志物 CD34 和 CD45 阴性。与 AD-MSCs 相比,EMSCs 的生长速度相似或略高,并且两种 EMSCs 和 AD-MSCs 的细胞比例相似,均处于 G0/G1 和 G2/M 期;然而,EMSCs 的细胞周期 S 期比例明显(P <.05)高于 AD-MSCs。通过实时定量聚合酶链反应(RT-qPCR)检测谱系特异性标记基因如 DES、ACTA2、cTnT 和 ACTC1 的表达,证实了分化细胞向心肌细胞样细胞的转分化能力。此外,完全分化的细胞表达心肌细胞特异性蛋白标记物 cTnT 和 ACTC1。通过形态学变化和β-细胞特异性标记基因如 PDX1、GLUT2、SST、NKX6.1、PAX4 和 NGN3 的表达分析,证实了 EMSCs 向胰腺β细胞样细胞的内胚层分化能力。经葡萄糖刺激后,分化细胞分泌胰岛素和 C 肽,免疫染色评估显示,细胞表达胰岛素、PDX1、PAX4、NGN3 和 GLUT2 等蛋白水平的标记物,证实了向β细胞样细胞的成功分化。猪 EMSCs 具有 MSCs 的所有特征,是研究细胞分化分子机制的合适模型。

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