中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (27): 4984-4987.doi: 10.3969/j.issn.1673-8225.2011.27.010

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

人脂肪源干细胞分离培养及向成骨细胞的诱导分化

白  雯1，尹  硕2，崔  磊2，江  明1   

  1. 1新疆医科大学第一附属医院血液科，新疆血液病研究所，新疆维吾尔自治区乌鲁木齐市  830054
    2上海组织工程研究与开发中心，上海市  200030
  • 收稿日期:2011-02-26 修回日期:2011-06-03 出版日期:2011-07-02 发布日期:2011-07-02
  • 通讯作者: 崔磊，博士，教授，上海组织工程研究与开发中心，上海市 200030 并列通讯作者：江明，硕士，主任医师，新疆医科大学第一附属医院血液科，新疆血液病研究所，新疆维吾尔自治区乌鲁木齐市 830054 jiangmingyy@yahoo.com.cn
  • 作者简介:白雯★，女，1981年生，新疆维吾尔自治区乌鲁木齐市人，汉族，2009年新疆医科大学毕业，硕士，医师，主要从事造血干细胞移植研究。 baiwenbox@yahoo.cn
  • 基金资助:

    研究生课题经费。

Isolation, culture and osteogenic differentiation of human adipose-derived stem cells 

Bai Wen1, Yin Shuo2, Cui Lei2, Jiang Ming1   

  1. 1Department of Hematology, First Affiliated Hospital of Xinjiang Medical University, Xinjiang Hematology Institute, Urumqi  830054, Xinjiang Uygur Autonomous Region, China
    2Shanghai Tissue Engineering Research and Development Center, Shanghai  200030, China
  • Received:2011-02-26 Revised:2011-06-03 Online:2011-07-02 Published:2011-07-02
  • Contact: Cui Lei, Doctor, Professor, Shanghai Tissue Engineering Research and Development Center, Shanghai 200030, China Correspondence to: Jiang Ming, Master, Chief physician, Department of Hematology, First Affiliated Hospital of Xinjiang Medical University, Xinjiang Hematology Institute, Urumqi 830054, Xinjiang Uygur Autonomous Region, China jiangmingyy@yahoo.com.cn
  • About author:Bai Wen★, Master, Physician, Department of Hematology, First Affiliated Hospital of Xinjiang Medical University, Xinjiang Hematology Institute, Urumqi 830054, Xinjiang Uygur Autonomous Region, China baiwenbox@yahoo.cn
  • Supported by:

    the Graduate Student Research Fund*

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被引次数

1

摘要：

背景：脂肪源干细胞可分泌众多的免疫调节因子，不引起T细胞的细胞毒作用，并可通过调整T淋巴细胞的种类和数量。
目的：探讨人脂肪源干细胞在体外分离培养扩增的方法及向成骨细胞诱导分化的能力。
方法：以0.1%的Ⅰ型胶原酶通过组织消化的方法分离人脂肪组织中的干细胞，体外扩增培养至第2代后检测其表面抗原的表达，并在成骨诱导液中促进其向成骨细胞的分化，通过碱性磷酸酶染色、茜素红染色及对碱性磷酸酶的RT-PCR检测来明确其分化能力。
结果与结论：体外分离培养的脂肪源干细胞生长稳定，扩增速度快。流式细胞仪检测结果显示其高表达干细胞相关抗原。向成骨细胞诱导后经免疫组化染色可见矿化结节形成，RT-PCR检测发现碱性磷酸酶表达阳性。提示脂肪源干细胞在体外分离培养方法简单，扩增速度快，并具有定向分化的能力，是可靠的组织修复和细胞治疗的种子细胞来源。

关键词: 脂肪源干细胞, 组织消化, 体外扩增, 成骨分化, 碱性磷酸酶

Abstract:

BACKGROUND: Adipose-derived stem cells (ADSCs) can secrete a large number of immune regulatory factors, not cause T-cell cytotoxicity, and adjust the type and quantity of T lymphocytes.
OBJECTIVE: To establish a method for the isolation and culture of human ADSCs and to explore the ability of osteogenic differentiation.
METHODS: ADSCs were isolated from the inguinal fat pads and digested with 0.1% type I collagenase, and cultured in osteogenic medium to study the osteogenic differentiation potential by using alkaline phosphatase staining, alizarin red staining and RT-PCR.
RESULTS AND CONCLUSION: ADSCs could be expanded steadily in vitro and flow cytometry revealed high expression of stem cell-associated markers. The cytoplasm of the induced cells became black by alkaline phosphatase staining, the mineralized nodule became reddish-brown by alizarin red, RT-PCR detection of alkaline phosphatase positive expression. Human ADSCs are an ideal source of adult stem cells for use in the organization repair and cell therapy.

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