恒温摇动法纯化培养大鼠视网膜Müller细胞的技术研究

A technique study on the purification culture of rat retinal Müller cells with constant-temperature shaking

目的 建立一种新的纯化Ｍ櫣ｌｌｅｒ细胞的技术方法。方法 将原代培养后的混合性视网膜细胞固定在一个摇床平台上 ,保持在 3 7℃。摇床转速调整到使培养液不生成泡沫的速度 ,过夜 (1 8～ 24ｈ)摇动后 ,将培养液上清弃去 ,通过免疫细胞化学的方法鉴定原代和摇动法得到的Ｍ櫣ｌｌｅｒ细胞的纯度。结果 原代培养的视网膜细胞有大量神经元和Ｍ櫣ｌｌｅｒ细胞混杂 ,摇动过程使得附着于Ｍ櫣ｌｌｅｒ细胞上的神经元细胞脱落 ,Ｍ櫣ｌｌｅｒ细胞的纯度通过ＧＦＡＰ确定可达 95%以上。结论 恒温摇床摇动的方法 。

ObjectiveTo establish a new method for the purification culture of retinal Müller cells in vitro.MethodPrimary confluent retinal mixed cell culture were taped to a rotary platform at 37℃.Setting platform rotate at a speed just below foaming of the medium.Platform was allowed shaking overnight(18～24 h),supernatant was removed.The primary and rotated-yielded Müller culture’s purification were identified by immuoncytochemistry.ResultMany neurons were mixed with Müller cells in the primary retinal cell.The shaking process allowed the neurons detach from Müller cells,the purification of Müller cells was approximately determined to 95% by GFAP.ConclusionThe constant-temperature shaking method provides a rapid and reliable method for purification of retinal Müller cells.