明胶对鸡胚盘细胞的增殖和多功能性的影响

[目的]本试验旨在解决鸡胚盘细胞（cBC）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（hVTN）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（EdU）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cBC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(cBC) 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(EdU) 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 cBC. And compared with the hVTN,treated by 10 g？L-1 gelatin on the wells has higher cost performance and higher application value