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不同来源的同种供体人沃顿氏胶间充质干细胞向功能性平滑肌样细胞分化的潜能。

Differentiation potential of different regions-derived same donor human Wharton's jelly mesenchymal stem cells into functional smooth muscle-like cells.

机构信息

Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 900 Gajwa, Jinju, 660-701, Republic of Korea.

Animal Biotechnology Division, National Institute of Animal Science, RDA, 1500 Kongjwipatjwi-ro Isero-myeon, Wanju-gun, Jeollabuk-do, 565-851, Republic of Korea.

出版信息

Cell Tissue Res. 2019 Aug;377(2):229-243. doi: 10.1007/s00441-019-03009-7. Epub 2019 Apr 3.

Abstract

The present study evaluates the transdifferentiation potential of different region-derived same donor Wharton's jelly MSCs (WJMSCs) into functional smooth muscle-like cells (SMLCs). All regions showed baseline expression for early smooth muscle cell (SMC) markers (αSMA and SM22-α) whereas mid marker CALPONIN gradually reduced during in vitro culture expansion and late marker myosin heavy chain type-11 (MHY-11) was completely absent. Furthermore, WJMSCs were induced to SMLCs using DMEM containing 10% FBS supplemented with different concentrations/combinations of TGF-β1 and PDGF-BB under normoxia (20% O) condition. Three treatment groups namely group A: 2.5 ng/ml TGF-β1, group B: 5 ng/ml PDGF-BB and group C: 2.5 ng/ml TGF-β1 + 5 ng/ml PDGF-BB were used for the induction of WJMSCs into SMLCs. Cells were evaluated for SMC-specific marker expression at different time intervals. Finally, selection of the SMC-specific highly potent region along with the most suitable treatment group was done on the basis of highest outcome in terms of SMC-specific marker expression and functional competence of transdifferentiated cells. Among all regions, baby region-derived WJMSCs (B-WJMSCs) exhibited highest SMC marker expression and functional ability. To mimic the in vivo physiological conditions, hypoxic conditions (3% O) were used to evaluate the effect of low oxygen on the SMLC differentiation potential of selected WJMSCs using previously used same parameters. Annexin-V assay was performed to check the effect of cytokines and different oxygen concentrations, which revealed no significant differences. It was concluded that different induction conditions have different but positive effects on the functional SMLC differentiation ability of WJMSCs.

摘要

本研究评估了不同来源同一供体的牙髓基质干细胞(WJMSCs)向功能性平滑肌样细胞(SMLCs)的转分化潜能。所有区域均表现出早期平滑肌细胞(SMC)标志物(αSMA 和 SM22-α)的基础表达,而中期标志物钙调蛋白在体外培养扩增过程中逐渐减少,晚期标志物肌球蛋白重链型-11(MHY-11)则完全缺失。此外,将 WJMSCs 在含 10%FBS 的 DMEM 中,在常氧(20%O)条件下,用不同浓度/组合的 TGF-β1 和 PDGF-BB 诱导为 SMLCs。三个处理组,即 A 组:2.5ng/mlTGF-β1、B 组:5ng/mlPDGF-BB 和 C 组:2.5ng/mlTGF-β1+5ng/mlPDGF-BB,用于诱导 WJMSCs 向 SMLCs 分化。在不同时间间隔评估细胞的 SMC 特异性标志物表达。最后,根据 SMC 特异性标志物表达和转分化细胞功能能力的最高结果,选择 SMC 特异性高潜力区域和最合适的处理组。在所有区域中,婴儿区来源的 WJMSCs(B-WJMSCs)表现出最高的 SMC 标志物表达和功能能力。为了模拟体内生理条件,使用低氧(3%O)来评估所选 WJMSCs 的 SMLC 分化潜能的影响,使用以前相同的参数。用 Annexin-V 检测细胞因子和不同氧浓度的影响,结果显示无显著差异。结论是,不同的诱导条件对 WJMSCs 功能性 SMLC 分化能力有不同但积极的影响。

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