凹凸界面对小鼠胚胎干细胞多能性的影响

Effects of Concave and Convex Interface on Pluripotency of Mouse Embryonic Stem Cells

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(alkaline phosphatase,ALP)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。凹面和凸面基底上干细胞的Oct4-GFP表达含量和ALP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。通过改变细胞外基质曲率,可以帮助胚胎干细胞体外培养维持多能性。研究结果对胚胎干细胞的研究和临床应用有重要意义。

Objective To explore the effect of concave and convex interface on 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 cell colony was observed.The pluripotency of embryonic stem cells was detected by immunofluorescence and alkaline phosphatase(ALP)staining.Results Embryonic stem cells on concave substrates and convex substrates had higher stereo degree and circularity than those on flat substrates,but it was more obvious on concave substrates.Besides,the expression level of Oct4-GFP and the staining intensity of ALP 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.Conclusions Compared with flat substrates,concave substrates and convex substrates had positive effects on the pluripotency maintenance of embryonic stem cells,which could help to maintain pluripotency,but concave substrates had better effects.Changing the substrate curvature could help to maintain pluripotency of embryonic stem cells cultured in vitro.The research findings are of great significance to the study and clinical application of embryonic stem cells.