Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran.
Iran Biomed J. 2021 Mar 1;25(2):99-105. doi: 10.29252/ibj.25.2.99.
Human embryonic stem cell-mesenchymal stem/stromal cell (hESCs-MSCs) open a new insight into future cell therapy applications, due to their unique characteristics, including immunomodulatory features, proliferation, and differentiation.
Herein, hESCs-MSCs were characterized by immunofluorescence technique with CD105 and FIBRONECTIN as markers and FIBRONECTIN, VIMENTIN, CD10, CD105, and CD14 genes using reverse transcription-polymerase chain reaction technique. Fluorescence-activated cell sorting was performed for CD44, CD73, CD90, and CD105 markers. Moreover, these fibroblast-like cells, due to multipotent characteristics, differentiated to the osteoblast.
MSCs were derived from diploid and triploid hESC lines using sequential three dimensional and two dimensional cultures and characterized with the specific markers. Immunofluorescence showed the expression of FIBRONECTIN and CD105 in hESCs-MSCs. Flow cytometry data indicated no significant difference in the expression of MSC markers after 6 and 13 passages. Interestingly, gene expression profiles revealed slight differences between MSCs from diploid and triploid hESCs. hESCs-MSCs displayed osteogenic differentiation capacity, which was confirmed by Alizarin red staining.
Our findings reveal that both diploid and triploid hESC lines are capable of forming MSCs; however, there are some differences in their gene expression profiles. Generation of MSCs from hESCs, as a non-invasive procedure in large scale, will lend itself for the future cell-based therapeutic applications.
人类胚胎干细胞-间充质干细胞（hESCs-MSCs）因其独特的特性，包括免疫调节特性、增殖和分化，为未来的细胞治疗应用开辟了新的视角。
在此，通过免疫荧光技术，使用 CD105 和纤维连接蛋白作为标记物，以及逆转录-聚合酶链反应技术，对 hESCs-MSCs 进行了鉴定。使用荧光激活细胞分选术对 CD44、CD73、CD90 和 CD105 标志物进行了检测。此外，这些成纤维样细胞由于具有多能性特征，可分化为成骨细胞。
通过顺序三维和二维培养，从二倍体和三倍体 hESC 系中获得了 MSC，并具有特定的标记物。免疫荧光显示 hESCs-MSCs 中纤维连接蛋白和 CD105 的表达。流式细胞术数据表明，在第 6 和 13 代后，MSC 标志物的表达没有显著差异。有趣的是，基因表达谱显示二倍体和三倍体 hESC 来源的 MSC 之间存在细微差异。hESCs-MSCs 表现出成骨分化能力，这通过茜素红染色得到了证实。
我们的研究结果表明，二倍体和三倍体 hESC 系均能够形成 MSC；然而，它们的基因表达谱存在一些差异。从 hESC 生成 MSC 是一种非侵入性的大规模操作，这将为未来的基于细胞的治疗应用提供帮助。
