1.Beijing Institute Of Fashion Technology;2.Beihang University;3.Tsinghua University
National Key R&D Projects
背景 体内细胞长期存在于动态的、复杂的微环境中，其中多种物理因素如几何结构、空间维度等作用于细胞，细胞能够感受这些物理因素的刺激并做出适当的响应。细胞外基质的物理性质对干细胞命运的影响受到越来越多的关注。目的 探究凹凸界面对体外培养的小鼠胚胎干细胞多能性维持的影响。方法 制作凹凸不同的细胞外基质培养小鼠胚胎干细胞，观察细胞的克隆形态，并通过免疫荧光和碱性磷酸酶染色，检测胚胎干细胞的多能性。结果 在凹面和凸面基底上胚胎干细胞的立体度和圆度均比平面基底高，但凹面基底更明显。并且凹面和凸面基底上干细胞的Oct4-GFP表达含量和碱性磷酸酶染色强度均明显高于平面基底，其中凹面基底更为显著。结论 与平面基底相比，凹面基底和凸面基底均对胚胎干细胞的多能性维持有积极影响，能够有助于维持全能性，但凹面基底效果更好。结果表明，通过改变细胞外基质的曲率可以帮助胚胎干细胞体外培养维持多能性，这对胚胎干细胞的研究和临床应用有重要意义。
Background The physical features of substrate have attracted more and more attention in the in vitro culture of stem cells. Objective To explore the effect of concave and convex interfaceon in vitro culture of mouse embryonic stem cells. Methods Mouse embryonic stem cells were cultured on substrate with concave and convex interface.. The biological morphology of the cell colony was observed. The pluripotency of embryonic stem cells was detected by immunofluorescence and alkaline phosphatase staining. Results Embryonic stem cells on concave substrates and convex substrates have higher stereo and circularity than those on flat substrates, but it is more obvious on concave substrates.Besides , the expression level of Oct4-GFP and the staining intensity of alkaline phosphatase in embryonic stem cells which were cultured on concave substrates and convex substrates were significantly higher than those on flat basement, especially on concave substrates. Conclusion Compared with flat substrates, concave substrates and convex substrates have positive effects on the pluripotency maintenance of embryonic stem cells, which can help to maintain pluripotency, but concave substrates have better effects. The results showed that changing the curvature of the substrate can help maintain pluripotency of embryonic stem cells cultured in vitro. It is of great significance to the research and clinical application of embryonic stem cells.