人羊膜上皮细胞的原代及传代培养_刘小勇.pdfVIP

2007 2 28 2 181 * 1 1 2 1 3 4 2 2 刘小勇 陈剑 吴静 周清 王彦平 李红 徐锦堂 赵松滨 1 2 3 4 暨南大学 附属第一医院眼科, 医学院眼科研究室, 病理生理学教研室, 附属第一医院病理科( 广州 510630) 建立体外培养人羊膜上皮 胞的方法, 观察体外培养的羊膜上皮 胞的生物学特性 取足月剖宫产术后羊膜, 经胶原酶和胰蛋白酶消化后, 获取的羊膜上皮 胞接种于含 10% 胎牛血清培养基中进 行原代和传代培养, 探索其合适的培养条件, 用倒置显微镜观察培养的人羊膜上皮 胞体外生长的特征用苏木 精-伊红染色扫描电镜和 胞角蛋白免疫组织化学染色的方法对培养 胞进行形态学观察和鉴定 人 羊膜上皮 胞可以在体外成功的培养传代,体外可连续传 8~ 10代体外培养 胞呈多角形,长满后呈上皮 胞 特有的铺路石样外观扫描电镜观察 胞表面有丰富的微绒毛 胞角蛋白 keratin 单克隆抗体染色阳性 人羊膜上皮 胞在体外可成功进行原代和传代培养, 体外培养的人羊膜上皮 胞在 一定时间内可维持增 殖能力 羊膜 上皮 胞 胞培养 The pr im ary cu lture and passage of hum an am n iotic ep ith elial ce lls LIUX iao- yong, CHEN Jian, WU Jing, et a.l TheD epartm ent of Ophthalm ology, The F irst ff ilia tedH osp ital of Jinan University, Guangzhou 510630, China A b stract O bj ective To estab lish am etho of culturing hum an am niotic ep ithelial cells in vitro an to stu y its b iologic features. M ethod s A p iece of amn ioticm embrane taken from an uncom plicate elective caesarean sectionw as -i geste by collagenase an tryps in respectively. The am niotic ep ithelial cells were see e in DMEM ( u lbeccos m o ifie eagles m e ium ) con taining 10% fetal bovine serum for prmi ary cu lture an passage. The optmi al cu lture con ition an feature of the am niotic epithelial cellsw ere stu ie in vitro. The cu lture cellsw ere investigate m orphologically byH em a- toxylin- eosin staining, scanning electron m icroscope( SEM ) an w ere i entifie by cytokeratin mi munoh istochem istry. R esu lts H um an amn iotic epithelial cellsw ere successfu lly cu lture an passage in vitro, an coul be passage succes- sively for 8- 10 tmi es. M ost of the culture cellsw ere polygon an of typ ical slabstone- like appearance. M anym icrovilli