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人类胚胎干细胞(hESC)的体外培养通常需要附着在蛋白质培养基质或饲养细胞层上,并主要依靠细胞的酶解离方法传代。采用动物来源的蛋白酶处理如此敏感的细胞会损伤膜蛋白、增加污染风险,这显然是不可取的,应尽量避免使用。在此,我们汇报了采用高通量方法筛选出一类以2-(二乙基氨基)乙基丙烯酸酯为主要单体的温敏型水凝胶。该类材料不仅具有在小幅度降温时改变性能以使其表面生长的干细胞脱粘附的功能,从而取代细胞传代中使用的生物酶,而且能取代蛋白质基质使hESC可以在体外进行长期培养(>20代),并保持完整的干细胞特征。这些具有可控化学结构的、可取代具动物来源生物基质的合成材料代表了一种新型的体系。它将是一种能使干细胞的培养达到可控、高效和安全的材料,会大大促进干细胞的深入研究和在临床医学中的应用。
In vitro culture of human embryonic stem cells (hESCs) usually requires attachment to protein culture media or feeder cell layers and passage of cells primarily by enzymatic dissociation methods. Treatment of such sensitive cells with animal-derived proteases can damage membrane proteins and increase the risk of contamination, which is clearly undesirable and should be avoided. Here, we report the use of high-throughput methods to screen out a class of 2- (diethylamino) ethyl acrylate as the main monomer of the temperature-sensitive hydrogel. This kind of material not only has the function of de-adhering stem cells whose surface properties are changed when the temperature is decreased slightly, but also replaces the biological enzyme used in cell passage, and can replace hESC for long-term culture in vitro (> 20 generations) and maintain intact stem cell characteristics. These synthetic materials that have a controlled chemical structure that can replace biological matrices of animal origin represent a new type of system. It will be a material that will make stem cell culture controllable, efficient and safe, which will greatly facilitate the further research of stem cells and their application in clinical medicine.