奶牛和绵羊子宫内膜细胞体外培养方法的研究

Study on the In-vitro Cultured Methods for the Uterine Endometrial Cells of Dairy Cattles and Sheep

为了探讨适用于奶牛和绵羊子宫内膜细胞的培养方法,采用了组织培养法、常温消化法、先组培后消化法和先消化后组培法,并适时用倒置显微镜观察,探讨哪一种原代培养方法更适合获得奶牛和绵羊子宫内膜细胞。结果表明,组织培养的奶牛和绵羊子宫内膜细胞生长较快,细胞形态为梭形和铺路石样交织在一起;常温消化法没有成功获得奶牛子宫内膜细胞,但获得了绵羊子宫内膜细胞;而先组培后消化法获得了奶牛子宫内膜细胞,但无法传代、不能长久生长;而先消化后组培法获得的细胞与单一组培法获得的奶牛和绵羊子宫内膜细胞情况相似,细胞上出现很多组织残块,覆盖在细胞上面不利于细胞增殖。说明组织培养方法是简捷快速获得牛、绵羊子宫内膜细胞的方法,获得的子宫内膜细胞可以在体外至少传4代,这类细胞可以用于研究动物子宫内膜的功能和相关分子调控机制。

In this experiment,it was aimed to investigate the suitable primary in-vitro cultured methods for the uterine endometrial cells of dairy cattles and sheep.Total four methods were compared with one another as following： the method by tissue culture only,the method by normal trypsin digestion only,the method by tissue cultured-trypsin digestion,and the one by trypsin digestion — tissue culture.The results showed that the uterine endometrial cells of dairy cattles and sheep took shape of either the spindle cell or the cobblestone morphology cell,normally grew faster than the method by tissue culture only than those by the other ones.The uterine endometrial cells of dairy cattles could not be obtained from the method by normal trypsin digestion only,but the uterine endometrial ones of sheep.Contrastly,through the method by the tissue cultured-trypsin digestion,the bovine uterine endometrial cells could be obtained,but could not be passaged and survival for a relatively long duration.In addition,through the method by the trypsin digestion-tissue culture,both kinds of uterine endometrial cells could be obtained similarly to the one by tissue culture only,but it easily displayed the residual block overlaying on the newly spreading cells,which would inhibit those viable cells from their proliferation.Collectively,it can be concluded that the method by tissue culture only is a shortcut,highly efficient way for establishing the in-vitro cultured model for the uterine endometrial cells of both dairy cattles and sheep.Such kind of uterine endometrial cells can be passaged for at least 4 generations,and be applied for the approriate analysis about their cellular function and molecular regulation.