鸡输卵管上皮细胞体外分离培养的研究

鸡输卵管上皮细胞是卵清蛋白的主要分泌细胞,是研究输卵管特异表达蛋白调控的重要工具。在以往的研究中,多采用普通DMEM培养液对鸡输卵管上皮细胞进行分离与培养,容易造成其自身特性在体外培养过程中的改变。本研究我们优化了细胞分离方法,发现从输卵管漏斗部组织分离的输卵管上皮细胞增殖较快;用鸡输卵管上皮细胞培养基相比DMEM更适合促进细胞生长;与胰酶相比,用Accutase消化酶进行细胞传代,有利于输卵管上皮细胞特性维持。对所获得的输卵管上皮细胞鉴定发现,己烯雌酚能促进卵清蛋白的表达,说明分离培养的细胞保持了鸡输卵管上皮细胞特性。本研究建立的方法为输卵管特异表达蛋白调控以及家禽生物反应器的研究奠定了基础。

Isolation and Culture of Chicken Oviduct Epithelial Cells in vitro

Chicken oviduct epithelial cells(OECs) are the main secretory cells of ovalbumin and useful tool for research in regulation of oviduct specific expressed protein. In previous studies, the chicken OECs was usually cultured in DMEM medium, which easily resulted in loss of OEC characteristics. In this study, we optimized the protocol for isolation of OECs and found that cells isolated from the infudibulum of the fallopian tube proliferated faster than those from the magnum. The OEC medium was more suitable for cell growth than cultured in DMEM. Compared to trypsin, accutase was beneficial for maintaining the characteristics of the OECs in continual subcultures.Incubation of the OECs in diethylstilbestrol revealed that the expression of ovalbumin was upregulated, indicating that the derived cells maintained the characteristics of chicken OECs. Thus, the protocol established facilitate the research in regulation of oviduct-specific protein expression and transgenic chicken for use as bioreactor.