摘要
目的探讨牛视网膜微血管内皮细胞(BREC)的体外选择性培养和鉴定方法。方法采用含0.075%胶原酶和0.03%牛血清白蛋白(BSA)的磷酸缓冲液(PBS)培养基选择性培养BREC。细胞鉴定采用经Ⅷ因子相关抗原免疫组化染色细胞鉴定内皮细胞。结果通过选择性培养获得的BREC纯度达98%以上,并能连续稳定传代。BREC早期聚集在微血管碎片周围,呈片状鹅卵石样单层生长,存在接触性抑制。经Ⅷ因子相关抗原免疫组化染色可见胞质内棕色的免疫沉积物反应呈阳性。结论本研究所采用的细胞培养方法可获得稳定生长与传代的较高纯度的视网膜微血管内皮细胞,简单、经济易操作。可为糖尿病视网膜病变等微血管病变研究提供可靠的(体外)试验方法。
Objective To probe a selective cultural method and the method to identify the bovine retinal capillary endothelial cells （BREC） in vitro. Methods With the isolation of active retinal blood vessels, and were cultured in a PBS medium supplemented with 0.03% bovine serum and 0.075% collagenase, BREC were identified by immunohistochemical method of Ⅷ related antigen. Results The purity of selectively cultured BREC was more than 98%, being reproducible. BREC got together around microvessel fragments with the small-cyprinoid-like configuration. After identified by immunohistochemical method of Ⅷ related antigen we could find positive reaction in cytochylema. Conclusion BREC with high purity can be easily and repeatably obtained, and identified by using slective culture with DMEM nutrient. Such anmial expermient afford a credible method for microangiopathy such as diabetic retinopathy.
出处
《世界临床药物》
CAS
2009年第7期411-413,共3页
World Clinical Drug
关键词
牛视网膜
微血管
内皮细胞
培养
鉴定
bovine retinal
capillary
endothelial cell
culture
identify
