中国组织工程研究

中国组织工程研究 ›› 2015, Vol. 19 ›› Issue (28): 4498-4502.doi: 10.3969/j.issn.2095-4344.2015.28.013

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

人脂肪干细胞的提取和鉴定

吴 尉1，梁 芳2，宋小琴2，胡平安2，刘 敏3   

  1. 1中南大学湘雅医学院附属肿瘤医院老干头颈内科，湖南省长沙市 410013；中南大学湘雅三医院，
    2内分泌科， 3营养科，湖南省长沙市 410013
  • 出版日期:2015-07-02 发布日期:2015-07-02
  • 通讯作者: 刘敏，中南大学湘雅三医院营养科，湖南省长沙市 410013
  • 作者简介:吴尉，男，1973年生，湖南省长沙市人，汉族，副主任医师，主要从事肺癌、干细胞治疗方面的研究。
  • 基金资助:

    湖南省科技厅项目(2013SK3060)

Extraction and identification of human adipose-derived stem cells 

Wu Wei1, Liang Fang2, Song Xiao-qin2, Hu Ping-an2, Liu Min3   

  1. 1Department of Head & Neck for Old Cadre, Tumor Hospital of Xiangya Medical College, Central South University, Changsha 410013, Hunan Province, China; 
    2Department of Endocrinology, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan Province, China; 3Department of Nutrition, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan Province, China
  • Online:2015-07-02 Published:2015-07-02
  • Contact: Liu Min, Department of Nutrition, Third Xiangya Hospital, Central South University, Changsha 410013, Hunan Province, China
  • About author:Wu Wei, Associate chief physician, Department of Head & Neck for Old Cadre, Tumor Hospital of Xiangya Medical College, Central South University, Changsha 410013, Hunan Province, China
  • Supported by:

     Project of Science and Technology Department of Hunan Province, No. 2013SK3060

PDF

638

被引次数

7

摘要：

背景：脂肪干细胞是存在于脂肪中的全能干细胞，具备自我更新能力与多向分化潜能，遗传背景相当稳定，体内植入后免疫排斥少，是一种比较理想的种子细胞。

目的：提取人大网膜脂肪干细胞，并进行成脂和成骨分化能力鉴定。
方法：收集手术患者大网膜的脂肪组织，经Ⅰ型胶原酶消化、过滤、离心后进行原代培养，观察细胞生长状态；用细胞计数法绘制人脂肪干细胞增殖曲线，计算处于对数生长期的倍增时间；用相应的定向诱导液诱导人脂肪干细胞向脂肪细胞及骨细胞方向分化，并用油红O染色、茜素红染色进行鉴定。

结果与结论：从手术患者大网膜脂肪组织中成功分离人脂肪干细胞，贴壁生长的人脂肪干细胞为长梭形，形态类似成纤维细胞。第3代人脂肪干细胞生长曲线呈S形，对数生长期人脂肪干细胞的倍增时间为45.90 h。人脂肪干细胞经成脂、成骨定向诱导分化2，3周后，油红O染色显示细胞内有大小不等的橘红色脂肪滴，茜素红染色可见着橘红色的典型钙化结节，提示所培养的脂肪干细胞具有向脂肪细胞、骨细胞系分化的能力。

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 干细胞, 脂肪干细胞, 诱导分化, 人脂肪干细胞, Ⅰ型胶原酶, 生长曲线, 脂肪细胞, 骨细胞, 大网膜, 油红O染色, 茜素红染色

Abstract:

BACKGROUND: Adipose-derived stem cells are totipotent stem cells in the adipose tissue, and have the function of self-renewal and multi-directional differentiation. Human adipose-derived stem cells are ideal seed cells with stable genetic milieu and few rejections.
OBJECTIVE: To extract human adipose-derived stem cells from human omental adipose tissue and to identify the cells by adipogenic and osteogenic induction.
METHODS: Omental adipose tissues were collected from surgical patients to isolate and culture adipose-derived stem cells using type I collagenase digestion, filtration and centrifugation. Cell growth was observed and proliferative curve of human adipose-derived stem cells were drawn by cell counting method to calculate the doubling time at logarithmic growth phase. After adipogenic and osteogenic induction, induced cells were identified using oil red O and alizarin red staining, respectively.
RESULTS AND CONCLUSION: Human adipose-derived stem cells were successfully isolated from the omentum tissues of surgical patients. Adherent cells were fusiform-shaped and like fibroblasts. The growth curve of passage 3 cells was in S shape, and the doubling time was 45.90 hours. After adipogenic and osteogenic induction for 2 and 3 hours, respectively, oil red O staining showed unequal-sized orange fat droplets, and alizarin red staining showed typical calcified nodules that were in orange. These findings indicate that adipose-derived .

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Adipose Tissue, Stem Cells, Collagenases, Cells, Cultured, Cell Differentiation

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