中国组织工程研究 ›› 2013, Vol. 17 ›› Issue (10): 1724-1729.doi: 10.3969/j.issn.2095-4344.2013.10.003

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

成人脂肪基质细胞体外扩增生长和超微结构

陆艳卉,元小冬,欧 亚,蔡亚楠,孙巧玉   

  1. 河北联合大学附属开滦总医院神经内科,河北省唐山市 063000
  • 收稿日期:2012-06-24 修回日期:2012-07-23 出版日期:2013-03-05 发布日期:2013-03-05
  • 通讯作者: 元小冬,教授,河北联合大学附属开滦总医院神经内科,河北省唐山市 063000 yxd68@sohu.com
  • 作者简介:陆艳卉★,女,1984年生,河北省唐山市人,汉族,2012年河北联合大学毕业,硕士,主要从事干细胞研究。 luyanhui012@163.com

In vitro amplification and ultrastructure of adult adipose-derived stromal cells

Lu Yan-hui, Yuan Xiao-dong, Ou Ya, Cai Ya-nan, Sun Qiao-yu   

  1. Department of Neurology, Affiliated Kailuan General Hospital of Hebei United University, Tangshan 063000, Hebei Province, China
  • Received:2012-06-24 Revised:2012-07-23 Online:2013-03-05 Published:2013-03-05
  • Contact: Yuan Xiao-dong, Professor, Department of Neurology, Affiliated Kailuan General Hospital of Hebei United University, Tangshan 063000, Hebei Province, China yxd68@sohu.com
  • About author:Lu Yan-hui★, Master, Department of Neurology, Affiliated Kailuan General Hospital of Hebei United University, Tangshan 063000, Hebei Province, China luyanhui012@163.com

摘要:

背景:脂肪基质细胞在体外能诱导分化为多种细胞,但脂肪基质细胞开始诱导反应的最佳时间尚未明确。
目的:通过成人脂肪基质细胞的体外扩增生长曲线和超微结构特征推测诱导分化反应的最佳时期。
方法:将取自吸脂术的脂肪组织消化、离心、提取细胞,并体外培养、传代。倒置相差显微镜下观察细胞形态学变化;细胞计数法测定脂肪基质细胞生长曲线;免疫组化和免疫荧光法检测CD44、CD29、CD34的表达;透射电镜观察脂肪基质细胞的超微结构。
结果与结论:脂肪基质细胞生长迅速,传代到第3代5-7 d呈对数生长,第8天细胞呈长梭形,数量达(5.32±0.03)×107 L-1并趋于稳定;脂肪基质细胞的CD44、CD29阳性表达率为(84.35±9.73)%,(86.37± 8.45)%,CD34为阴性表达;透射电镜观察到处于幼稚状态的细胞器等超微结构。脂肪基质细胞作为一种多能干细胞,传至第3-6代8-12 d时的细胞数量多且形态稳定,具有细胞分化和合成所需蛋白的超微结构,此时是开始诱导分化反应的最佳时期。

关键词: 干细胞, 脂肪干细胞, 脂肪基质细胞, 体外培养, 鉴定, 超微结构, 形态学特征, 生长曲线, CD44, CD29, 干细胞图片文章

Abstract:

BACKGROUND: Adipose-derived stromal cells can be induced to differentiate into various cells in vitro. However, the best induced time of adipose-derived stromal cells has not been well understood.
OBJECTIVE: To speculate the best period for induced differentiation response through in vitro amplification curve and ultrastructure of adult adipose-derived stromal cells.
METHODS: Adipose tissue from liposuction surgery was digested and centrifuged. Cells were extracted, cultured and passaged in vitro. Morphological changes of cells were observed under the inverted phase contrast microscope; the growth curve of adipose-derived stromal cells was drawn by cell count method; immunohistochemistry and immunofluorescence staining were used to detect CD44, CD29 and CD34 expression; and the ultrastructure of adult adipose-derived stromal cells was observed under transmission electron microscope.
RESULTS AND CONCLUSION: Adipose-derived stromal cells grew rapidly. Cells displayed logarithmic growth at passage 3 after cultured for 5-7 days, and the number of which reached (5.32±0.03) ×107/L after cultured for 8 days, then was stably kept, and the cells were in long-spindled shape. The positive rate of CD29 and CD44 was (84.35±9.73)% and (86.37±8.45)% respectively, whereas CD34 expression was negative by immunohistochemistry and immunofluorescence staining. Naive organelles were observed under the transmission electron microscope. As multiple stem cells, adipose-derived stromal cells, at passage 3 and 6 after cultured for 8-12 days, were most in number and stable in the shape and had the ultrastructures which enable cell differentiation and synthesize the required protein. This time is the best time for induced differentiation response.

Key words: stem cells, adipose-derived stem cells, adult adipose-derived stromal cells, in vitro culture, identification, ultrastructure, morphological characteristics, growth curve, CD44, CD29, stem cell photographs-containing paper

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