中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (14): 2512-2516.doi: 10.3969/j.issn.1673-8225.2011.14.010

• 脂肪干细胞 adipose-derived stem cells • 上一篇    下一篇

大鼠腹股沟脂肪间充质干细胞的分离、培养、鉴定及活体标记

毕晓娟1，马  艳1，江  明2   

  1. 新疆医科大学第一附属医院，1医学研究中心干细胞研究室，2血液二科，新疆维吾尔自治区乌鲁木齐市  830054
  • 收稿日期:2010-11-08 修回日期:2011-03-01 出版日期:2011-04-02 发布日期:2013-11-02
  • 通讯作者: 江明，硕士，教授，新疆医科大学第一附属医院血液二科，新疆维吾尔自治区乌鲁木齐市 830054 jiangmingyy@yahoo.com.cn
  • 作者简介:毕晓娟，女，1983 年生，新疆维吾尔自治区乌鲁木齐市人，汉族，2007 年新疆医科大学毕业，实习研究员，主要从事干细胞方面的研究。 Aibi1983@yahoo.com.cn
  • 基金资助:

    新疆维吾尔自治区自然科学基金项目(200821113)，项目名称：脂肪源干细胞对大鼠的造血重建功能及重建后免疫调节功能影响的作用研究。

Isolation, culture, identification and in vivo marker of inguinal adipose mesenchymal stem cells in rats

Bi Xiao-juan1, Ma Yan1, Jiang Ming2   

  1. 1Stem Cell Institute, Medical Research Center, the First Affiliated Hospital of Xinjiang Medical University, Urumqi  830054, Xinjiang Uygur Autonomous Region, China
    2Second Department of Blood, the First Affiliated Hospital of Xinjiang Medical University, Urumqi  830054, Xinjiang Uygur Autonomous Region, China
  • Received:2010-11-08 Revised:2011-03-01 Online:2011-04-02 Published:2013-11-02
  • Contact: Jiang Ming, Master, Professor, Second Department of Blood, the First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China jiangmingyy@yahoo.com.cn
  • About author:Bi Xiao-juan, Internship, Stem Cell Institute, Medical Research Center, the First Affiliated Hospital of Xinjiang Medical University, Urumqi 830000, Xinjiang Uygur Autonomous Region, China Aibi1983@yahoo.com.cn
  • Supported by:

    the Natural Science Foundation of Xinjiang Uygur Autonomous Region, No. 200821113*

PDF

542

被引次数

18

摘要：

背景：脂肪间充质干细胞具有来源丰富、取材方便、体外有较强增殖能力并具有多向分化的特点，有望成为骨组织工程、细胞治疗等的种子细胞。
目的：培养扩增大鼠脂肪源间充质干细胞，以活体标记并鉴定其分化潜能。
方法：无菌条件下取大鼠双侧腹股沟脂肪，Ⅰ胶原酶消化法分离培养脂肪源性干细胞，胰蛋白酶消化法传代扩增。取第3代脂肪干细胞进行流式检测HCAM、CD106、CD29、CD49D和CD34，生长曲线测定，吉姆萨染色，菲立磁标记，成脂诱导后油红O染色及成骨诱导后茜素红染色钙结节。
结果与结论：细胞呈长梭形漩涡样生长，第3代细胞流式鉴定CD29阳性，CD34、HCAM、CD49d、CD106低表达，生长曲线测定有对数生长期，平台期，菲立磁标记阳性率达80%，并且在一些诱导剂下分化为脂肪细胞及成骨细胞。提示，来源于大鼠腹股沟脂肪分离获得的脂肪源干细胞易于培养和传代扩增，并可活体标记且在特殊条件下可分化为成骨细胞和脂肪细胞。

关键词: 脂肪干细胞, 成脂分化, 成骨分化, 流式鉴定, 菲立磁标记

Abstract:

BACKGROUND: Adipose-derived stem cells (ADSCs) are easily collected and abundantly cultured, ADSCs can proliferate rapidly when being cultured in vitro, with multi-directional differentiation potential, ADSCs are expected as seed cells for tissue engineering and cell therapy.
OBJECTIVE: To culture rat ADSCs in vitro, and to mark and identify its differentiation.
METHODS: CollagenaseⅠdigestion was used to isolate ADSCs from rat groin fat pads under sterile condition. ADSCs were passaged and amplified by the trypsin digestion. At the third passage, growth curve of the third passage ADMSCs was drawn by cytometry. Cellular surface antigens HCAM, CD106, CD29, CD49D and CD34 were examined using flow cytometry. Giemsa staining, Feridex-labeled and induced to differentiate into osteoblasts and lipocytes. Oil red staining was used to examine lipocyte induction, alizarin red staining wwas used to examine osteoblast induction.
RESULTS AND CONCLUSION: In vitro cultured ADSCs were mainly spindle-shaped and whirlpool-shaped arranged. The third passage ADSCs were positive for CD29, but negative for CD34, HCAM, CD49d and CD106. Curve of growth detection had logarithmic phase and platform phase, the positive expression of Feridex reached 80%. ADMSCs were differentiated into lipocytes and osteoblasts under certain conditions. ADSCs isolated from rat epididymal fat pads can be easily cultured, passaged and amplified, and which can mark alive and differentiate into osteoblasts and adipocytes under certain conditions.

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