明胶对鸡胚盘细胞的增殖和多功能性的影响

Effects of gelatin on proliferation and pluripotency of chicken blastodermal cells

[目的]本试验旨在解决鸡胚盘细胞(c BC)不易贴壁的问题及应用E8培养基建立新的鸡胚盘细胞培养体系。[方法]在E8培养基基础上,应用10 g·L^(-1)明胶和人重组玻连蛋白(h VTN)包被培养板,对比其在促进鸡胚盘细胞贴壁速度、促进增殖和维持多功能性方面的差异。[结果]使用10 g·L^(-1)明胶包被培养板后,鸡胚盘细胞12 h内即可迅速贴壁。10 g·L^(-1)明胶处理组中5-乙炔基-2'-脱氧尿嘧啶核苷(Ed U)阳性细胞率显著高于对照组(P<0.01),并且能够抑制凋亡基因P21、Fas的mRNA表达(P<0.05),进而促进细胞增殖,并且能够促进多功能基因Nanog、PouV的表达来维持c BC的多功能性。[结论]10 g·L^(-1)明胶可以作为一种新的包被试剂来进行cBC的培养,与人重组玻连蛋白相比,明胶体现了更高的性价比,具有较高的应用价值。

[Objectives]This paper aims to overcome the difficulties of adherence of chicken blastodermal cells（c BC）and to establish a new culture system for chicken blastodermal cells by using E8 medium.[Methods]By using 10 g·L^-1 gelatin-coated and human recombinant vitronectin（hVTN）-coated wells,we compared with each other in the adherent rate and maintaining pluripotency of chicken blastodermal cells.[Results]Treated by 10 g·L^-1 gelatin on the wells,chicken blastodermal cells could be pasted to the wells within 12 hours.5-ethynyl-2＇-deox-youridine（Ed U） positive cells in the 10 g·L^-1 gelatin treatment group were significantly higher than those in the control（P〈0.01）,and could inhibit the mRNA expression of P21 and Fas（P 0.05）,and thus promote cell proliferation,and promote the expression of multifunctional genes（Nanog,PouV） to maintain pluripotency of cBC.[Conclusions]10 g·L^-1 gelatin can be used as a new embedding reagent for culturing c BC.And compared with the hVTN,treated by 10 g·L^-1 gelatin on the wells has higher cost performance and higher application value.