人食管癌间质干样细胞分离培养及生物学特性鉴定

基础医学与临床 ›› 2012, Vol. 32 ›› Issue (7): 798-803.

人食管癌间质干样细胞分离培养及生物学特性鉴定

周中卫¹,许文荣²,施舜缤³,胡嘉波¹,朱孝中³,王晓慧

收稿日期:2011-08-22 修回日期:2011-12-01 出版日期:2012-07-05 发布日期:2012-06-20
通讯作者: 胡嘉波 E-mail:hu@ujs.edu.cn
基金资助:
江苏大学高级人才专项资助;江苏大学临床医学科技发展基金

Isolation of mesenchymal stem-like cells from human esophageal carcinoma and identification of their biological characteristics

Received:2011-08-22 Revised:2011-12-01 Online:2012-07-05 Published:2012-06-20

摘要/Abstract

摘要： 摘要：目的探讨人食管癌间质干样细胞（mesenchymal stem-like cells from human esophageal carcinoma，hEC-MSCs）体外分离培养方法，并对其生物学特性进行鉴定。方法用胶原酶消化法分离培养人食管癌间质干样细胞，RT-PCR 检测基因表达，流式细胞仪检测表面标记和细胞周期，绘制生长曲线，检测染色体核型，成骨诱导和心肌诱导检测其多向分化潜能。结果胶原酶消化法分离的hEC-MSCs呈长梭形样，细胞增殖迅速，表达间质干细胞（MSCs）相关基因Oct-4、Nanog、vimentin、N-cadherin，不表达上皮细胞相关基因E-cadherin；表达MSCs相关表面标记CD13、CD29、CD44、CD105，不表达CD33、CD45、CD133、CD14、CD34及HLA-DR。细胞周期分析显示多数细胞处于G0/G1期，少数在S和G2/M期。细胞内含有 46 条染色体, 形态正常；具有分化为成骨细胞及心肌细胞的潜能。结论胶原酶消化法能有效分离hEC-MSCs，对进一步研究食管癌的微环境具有重要意义。

关键词: 关键词：食管癌, 间质干样细胞, 胶原酶, 鉴定

Abstract: Abstract Objective To isolate mesenchymal stem-like cells from human esophageal carcinoma and to identify their biological characteristics. Methods hEC-MSCs were isolated by collagenase digestion.Gene expression were detected by RT-PCR; surface antigen markers and cell cycle were determined by flow cytometry. Growth curve was portrayed and chromosme karyotype was assayed.Multilineage differentiation capacity of the cells was tested by inducing differentiation toward osteoblasts and cardiomyocytes. Results hEC-MSCs isolated by collagenase tended to be long and stringy in shape and multiplied rapidly.The cells expressed related gene Oct-4,Nanog,vimentin and N-cadherin,but not E-cadherin；FCM results showed that The cells expressed CD13, CD29,CD44 and CD105,but did not express CD33,CD45, CD133,CD14,CD34 and HLA-DR.Cell cycle analysis revealed that the majority of cells stood in G0/G1 phase while a small population of cells were in S and G2/M phase.The cells contain 46 chromosomes with normal Karyotype and they have the potentials differentiated into osteoblasts and cardiomyocytes.Conclusion hEC-MSCs can be isolated efficiently by collagenase digestion and they have important significance for further studying the microenvironment of esophageal carcinoma.

Key words: Key words esophageal carcinoma, hEC-MSCs, collagenase, identification

中图分类号:

中图分类号：R730.23

周中卫许文荣施舜缤胡嘉波朱孝中王晓慧. 人食管癌间质干样细胞分离培养及生物学特性鉴定[J]. 基础医学与临床, 2012, 32(7): 798-803.

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