【摘要】 目的研究Wistar大鼠胰岛细胞分离、纯化和培养的改良方法和条件,为进行细胞电生理实验提供胞膜完整、活性良好的胰岛细胞。方法水合氯醛腹腔麻醉,采用胶原酶ⅴ逆行灌注、原位消化及Ficoll-400梯度离心的方法分离、纯化大鼠胰岛,并分离、培养胰岛单细胞。结果采用该方法分离、纯化胰岛的获得率稳定;急性分离、培养的胰岛细胞12h～24h贴壁,(4～5)d胰岛细胞生长状态最佳,胞膜完整,折光性好。结论逆行灌注、原位消化及Ficoll-400梯度离心为一种有效的分离、纯化胰岛细胞的方法,培养的胰岛细胞(4～5)d达到最佳状态,适合进一步研究的要求。更多还原

【Abstract】 Objective To study on the optimal method and condition of the isolation,purification and culturing of wistar rat islet cells,so as to supply islet cells with complete membrane and fine activity,for researching on the cellular electro-physiology.Methods After anesthetizing rat intraperitoneal with chloraldurat,isolate and purify rat islet adopting CollagenⅤ retroperfusion,situ digestion and gradient centrifugation in Ficoll-400 solution.Then isolating and cultivating single islet cells.Results This method may gain steady yield of pancreatic islet.The isolation and cultivation of islet cells:adhering within 12～24 hours,optimal cells growing with complete membrane and fine refraction at 4～5 days.Conclution Retro perfusion,situ digestion and gradient centrifugation in Ficoll-400 solution is an effective method to isolate and purify islet cells of rats.The cultured islet cells have the optimal state at 4～5 days,fitting the further study.更多还原