Department of Pediatric Surgery, Faculty of Medicine, Gaziosmanpasa University, 60100 Tokat, Turkey.
Department of Medical Genetics, Yeditepe University Medical School, 34755 Istanbul, Turkey; Department of Biotechnology, Institute of Science, Yeditepe University, 34755 Istanbul, Turkey.
Exp Mol Pathol. 2018 Dec;105(3):395-403. doi: 10.1016/j.yexmp.2018.11.003. Epub 2018 Nov 8.
Amniotic fluid (AF) is the liquid layer that provides mechanical support and allows movement of the fetus during embryogenesis. Mesenchymal stem cells (MSCs), which have differentiation capacity, are also found in AF-derived cells at a low ratio. Smooth muscle cells (SMCs) play an important role in organ function and are frequently used in tissue engineering. We examined the differentiation of AF-derived MSCs (AMSCs) into SMCs. AMSCs were sorted from cultured amniotic cells and differentiated into SMCs using differentiation agents, including platelet-derived growth factor BB (PDGF-BB) and tumor growth factor β (TGF-β). Characterization of differentiated SMCs was confirmed morphologically, molecularly (via quantitative polymerase chain reaction [qPCR] and immunocytochemistry [ICC]), and functionally (using a contractile assay and fluo-4 calcium signaling assay). Poly(lactide-co-glycolide) (PLGA) scaffolds were fabricated, and the attachment capacity of AMSCs was assessed via scanning electron microscopy. AMSCs were successfully differentiated into SMCs. Our results indicate that AMSCs change their morphology and exhibit increased expression of ACTA2 and MYH11, which was confirmed via qPCR and ICC. Furthermore, functional experiments revealed that differentiated SMCs had both contraction ability and increased Ca concentration in the cytoplasm. Finally, PLGA scaffolds were prepared and AMSCs were successfully planted onto the scaffolds. The AMSCs fully differentiated into functional SMCs, and the PLGA polymer is a suitable scaffold material for AMSCs. With further clinical trials, AF-derived MSC-based SMC engineering may become a highly efficient treatment option.
羊水（AF）是在胚胎发生过程中为胎儿提供机械支撑和允许运动的液体层。间充质干细胞（MSCs）也具有分化能力，在 AF 衍生细胞中以低比例存在。平滑肌细胞（SMCs）在器官功能中起着重要作用，并且经常用于组织工程。我们研究了 AF 衍生的间充质干细胞（AMSCs）向 SMCs 的分化。从培养的羊膜细胞中分离出 AMSCs，并用分化剂将其分化为 SMCs，包括血小板衍生生长因子 BB（PDGF-BB）和转化生长因子 β（TGF-β）。分化的 SMCs 的特征通过形态学、分子学（通过定量聚合酶链反应[qPCR]和免疫细胞化学[ICC]）和功能学（使用收缩测定和 fluo-4 钙信号测定）进行了确认。制备了聚（乳酸-共-乙醇酸）（PLGA）支架，并通过扫描电子显微镜评估了 AMSCs 的附着能力。AMSCs 成功分化为 SMCs。我们的结果表明，AMSCs 改变其形态，并表现出 ACTA2 和 MYH11 的表达增加，这通过 qPCR 和 ICC 得到了证实。此外，功能实验表明，分化的 SMCs 具有收缩能力和细胞质中 Ca 浓度的增加。最后，制备了 PLGA 支架，并成功地将 AMSCs 种植在支架上。AMSCs 完全分化为功能性 SMCs，PLGA 聚合物是 AMSCs 的合适支架材料。随着进一步的临床试验，基于 AF 衍生 MSC 的 SMC 工程可能成为一种高效的治疗选择。
