摘要
目的建立人眼脉络膜血管内皮细胞快速培养方法并观察细胞的特征，为研究与人眼脉络膜血管内皮细胞有的关眼部疾病提供体外研究模型。方法采用胰酶、胶原酶对人眼脉络膜组织块进行两步消化，应用免疫磁珠CD31 Dynabeads对消化后的细胞混合悬液分选纯化后获得人眼脉络膜血管内皮细胞并进行细胞培养；采用形态学观察、透射电子显微镜扫描、内皮细胞特异标志F因子、CD31、CD34的免疫组织化学染色观察培养的人眼脉络膜血管内皮细胞特征。结果培养的细胞呈多边形、类圆形，细胞融合后呈铺路石样外观，传代后的细胞仍保持圆形或多边形，融合成单层时呈鹅卵石样排列，细胞表面吸附的磁珠明显减少；培养细胞胞浆内有内皮细胞特异性的棒状小体（Weibel-Palade小体），内皮细胞特异标志F因子、CD31、CD34染色阳性。结论本实验研究成功培养出人眼脉络膜血管内皮细胞，纯度较高，方法易行，可获得足够实验研究的细胞数量。
Objective To establish a rapid in vitro culture method of human choroidal endothelial cells （HCEC） and the cellular Characteristics to provide an in vitro model for researches of choroiretinal diseases which involved the HCEC. Methods The human choroidal tissues were digested in two steps by trypsin and collagenase, and the HCEC were obtained and cultured after the digested cell suspension was sorted and purified with magnetic beads of CD31 Dynabeads. The characteristics of HCMEC were observed by the morphologic observation method, transmission electron microscopy, and immunohistochemical staining with FⅧ factor, CD31, and CD34. Results The cultured HCEC were polygonal and oval, and after amalgamation, the cells had slabstone-like appearance. After the subculture, the configuration of HCEC remained the same, and represented cobblestone appearance with less magnetic beads attached on the cellular surface after HCEC converged into a single layer. The Weibel-Palade body which is the characteristic marker of endothelial cells was found. The staining of FⅧ fatcor, CD31, CD34 were positive. Conclusion HCEC can be cultured in vitro successfully with our method, which is easy to get sufficient number of highly purified HCEC.
出处
《中华眼底病杂志》
CAS
CSCD
北大核心
2007年第2期126-129,共4页
Chinese Journal of Ocular Fundus Diseases
