中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (46): 8665-8667.doi: 10.3969/j.issn.1673-8225.2011.46.027

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

小鼠睾丸间质细胞体外原代培养方法的建立

赵为民1，杨建英2，代  涛1，张功宝3，袁德品3   

  1. 1河南科技大学第三附属医院东方医院，河南省洛阳市 471003
    2河南科技大学医学技术与工程学院，河南省洛阳市 471003
    3中国平煤神马医疗集团总医院整形烧伤科，河南省平顶山市  467000
  • 收稿日期:2011-04-18 修回日期:2011-05-13 出版日期:2011-11-12 发布日期:2011-11-12
  • 通讯作者: 杨建英，博士，副教授，河南科技大学医学技术与工程学院，河南省洛阳市 471003 jyyang@yahoo.cn
  • 作者简介:赵为民，男，1957年生，上海市人，汉族，副主任医师，主要从事干细胞生物学与组织修复的研究。 jyyang2010@163.com
  • 基金资助:

    河南省科技计划项目(112102310144)，河南省教育厅自然科学研究计划项目(2011B310004)，河南科技大学博士启动基金项目(09001285)。

Establishment of a method for primary culture of mouse Leydig cells in vitro

Zhao Wei-min1, Yang Jian-ying2, Dai Tao1, Zhang Gong-bao3, Yuan De-pin3   

  1. 1Luoyang Dongfang Hospital, Third Affiliated Hospital of Henan University of Science and Technology, Luoyang 471003, Henan Provice China
    2College of Medical Technology and Engineering, Henan University of Science and Technology, Luoyang  471003, Henan Provice  China
    3Department of Burn and Plastic Surgery, China Pingmei Shenma Medical Group General Hospital, Pingdingshan  467000, Henan Province China
  • Received:2011-04-18 Revised:2011-05-13 Online:2011-11-12 Published:2011-11-12
  • Contact: Yang Jian-ying, Doctor, Associate professor, College of Medical Technology and Engineering, Henan University of Science and Technology, Luoyang 471003, Henan Province China jyyang@yahoo.cn
  • About author:Zhao Wei-min, Associate chief physician, Luoyang Dongfang Hospital, Third Affiliated Hospital of Henan University of Science and Technology, Luoyang 471003, Henan Province China jyyang2010@163.com
  • Supported by:

    Science and Technology Development of Henan Province, No. 112102310144*; the Natural Science Research Development Program of Education Department of Henan Province, No. 2011B310004*; Ph.D. Programs of Henan University of Science and Technology, No. 09001285*

PDF

585

被引次数

13

摘要：

背景：组织块培养睾丸生殖细胞对污染不容易控制，胰蛋白酶消化接种培养睾丸生殖细胞可能损伤细胞。
目的：建立一种切实可行的小鼠睾丸间质细胞体外原代培养方法。
方法：小鼠睾丸间质细胞的分离采用胶原酶消化法，纯化采用Percoll等密度梯度离心法，活率鉴定采用锥虫蓝拒染法，纯度鉴定采用3β-羟基固醇脱氢酶染色方法。
结果与结论：小鼠睾丸间质细胞纯度达可达90%以上；体外培养的细胞形态完整、增殖速度快、贴壁生长状态良好。说明实验成功建立了小鼠睾丸间质细胞的体外原代培养模型。

关键词: 睾丸间质细胞, 小鼠, 分离, 原代培养, 体外

Abstract:

BACKGROUND: Tissue explant culture of Leydig cells leads to difficult control of contamitation and trypsin digestion of Leydig cells may injury cells.
OBJECTIVE: To establish a simple and effective method for primary culture of mouse Leydig cells.
METHODS: Mature mouse Leydig cells were digested by collagenase digestion and were purified by Percoll gradient centrifugation. Cell viability was identified by Trypan blau staining and cell purity was determined by 3 β-hydroxysteroid dehydrogenase staining.
RESULTS AND CONCLUSION: The cell purity was > 90%. The Leydig cells retained complete morphology, raid proliferation and good adherence. Results showed that an in vitro primary culture model of mouse Leydig cells was successfully established.

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