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人羊膜上皮细胞的原代及传代培养
  • ISSN号:1001-9448
  • 期刊名称:《广东医学》
  • 时间:0
  • 分类:R329.25[医药卫生—人体解剖和组织胚胎学;医药卫生—基础医学]
  • 作者机构:[1]暨南大学附属第一医院眼科,广州510630, [2]暨南大学医学院眼科研究室,广州510630, [3]暨南大学病理生理学教研室,广州510630, [4]暨南大学附属第一医院病理科,广州510630
  • 相关基金:广东省重大科技计划项目(编号:2003A3020104),国务院侨办重点学科基金项目(编号:51205004)
中文摘要:

目的建立体外培养人羊膜上皮细胞的方法,观察体外培养的羊膜上皮细胞的生物学特性。方法取足月剖宫产术后羊膜,经胶原酶和胰蛋白酶消化后,获取的羊膜上皮细胞接种于含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细胞体外生长的特征。用苏木精-伊红染色、扫描电镜和细胞角蛋白免疫组织化学染色的方法对培养细胞进行形态学观察和鉴定。结果人羊膜上皮细胞可以在体外成功的培养传代,体外可连续传8~10代。体外培养细胞呈多角形,长满后呈上皮细胞特有的铺路石样外观。扫描电镜观察细胞表面有丰富的微绒毛。细胞角蛋白keratin单克隆抗体染色阳性。结论人羊膜上皮细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细胞在一定时间内可维持增殖能力。

英文摘要:

Objective To establish a method of culturing human amniotic epithelial cells in vitro and to study its biologic features. Methods A piece of amniotic membrane taken from an uncomplicated elective caesarean section was digested by collagenase and trypsin respectively. The amniotic epithelial cells were seeded in DMEM (dulbecco's modified eagle's medium) containing 10% fetal bovine serum for primary culture and passage. The optimal culture condition and feature of the amniotic epithelial cells were studied in vitro. The cultured cells were investigated morphologically by Hematoxylin- eosin staining, scanning electron microscope (SEM) and were identified by cytokeratin immunohistochemistry. Results Human amniotic epithelial cells were successfully cultured and passaged in vitro, and could be passaged successively for 8 - 10 times. Most of the cultured cells were polygon and of typical slabstone - like appearance. Many microvilli were observed on cell sur faces by SEM. Cytokeratin staining was positive. Conclusion Human amniotic epithelial cells can be successfully primary cultured and passaged in vitro, and the cells can proliferate for a certain period.

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期刊信息
  • 《广东医学》
  • 北大核心期刊(2011版)
  • 主管单位:广东省卫生和计划生育委员会
  • 主办单位:广东省医学学术交流中心
  • 主编:苏焕群
  • 地址:广州市惠福西路进步里2号之6
  • 邮编:510180
  • 邮箱:gdyx1963@163.com zhuangxw@126.com
  • 电话:020-81906074 81865066
  • 国际标准刊号:ISSN:1001-9448
  • 国内统一刊号:ISSN:44-1192/R
  • 邮发代号:46-66
  • 获奖情况:
  • 中国科技论文统计源期刊,美国《化学文摘》(CA)收录期刊
  • 国内外数据库收录:
  • 美国化学文摘(网络版),日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:70380