大鼠脉络丛上皮细胞的原代培养与鉴定

目的建立一种简便、实用的大鼠脉络丛上皮细胞的原代培养方法,以便进一步对其进行体外实验研究。方法无菌条件下取1～9d的Sprague-Dawley大鼠脉络丛组织,经机械分离后将细胞种植于含100mL/L胎牛血清的DMEM培养基中进行培养,定期进行形态学观察并摄片。在培养第9天左右,采用电子显微镜及Hematoxylin&Eosin(HE)染色进行形态学观察,免疫细胞化学染色法鉴定并计算脉络丛上皮细胞的纯度。结果培养的脉络丛上皮细胞呈梭形或多角形,电子显微镜下有微绒毛,细胞纯度在95%以上。结论利用机械分散法分离新生大鼠脉络丛上皮细胞,进行原代培养可获得高纯度的脉络丛上皮细胞,该方法简便实用。

Primary culture and identification of choroid plexus epithelial cells from neonatal rats

Objective To establish a simple but practical experiment method or procedure for primary culture of choroid plexus epithelial cells(CPECs) from rats so as to be used for further in vitro experiments. Methods CPECs were dissociated mechanically from neonatal Sprague-Dawley rats(1-9 days old),and cultured in DMEM nutrient with 100 mL/L fetal calf serum in water-saturated environment of 50 mL/L CO2 at 37 ℃.The morphological changes of cultured CPECs were observed.Hematoxylin and eosin(HE) staining,electron microscopy,and immunocytochemical technique were used for CPECs identification on day 9 of culture;the purity of positive cells was calculated. Results The cultured CPECs showed fusiform or multipolar morphology,or frequently clustered in irregular folds or were covered with irregular and numerous microvilli observed under electron microscope.Over 95% of the cultured cells were identified as CPECs or TTR-positive cells. Conclusion CPEC with high purity has been obtained in vitro from neonatal rats by mechanical dissociation without enzyme digestion,which is a simple and inexpensive way of primary cell culture.