上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2019, Vol. 39 ›› Issue (11): 1226-.doi: 10.3969/j.issn.1674-8115.2019.11.002

• 论著·基础研究 • 上一篇    下一篇

2类小鼠胎肝基质细胞的生物学特征及表达谱分析

王沄汉1，迟亚男1，杨冠恒2，范书玥1，马 姬1，薛 燕1, 2，曾凡一1, 2   

  1. 1. 上海交通大学基础医学院组织胚胎学与遗传发育学系，上海 200025；2. 上海交通大学附属儿童医院上海医学遗传研究所，国家卫生和健康委员会医学胚胎分子生物学重点实验室，上海市胚胎与生殖工程重点实验室，上海 200040
  • 出版日期:2019-11-28 发布日期:2019-12-16
  • 作者简介:王沄汉（1991—），男，硕士生；电子信箱：wyhauannng@163.com。
  • 基金资助:
    国家重点基础研究发展计划（2014CB964701，2014CB964703）；国家自然科学基金（31871484）；上海市“重中之重”临床重点学科建设项目（2017ZZ02019）；上海市自然科学基金（16ZR1428600）

Biological characteristics and gene profile analysis of two kinds of mofetal liver stromal cells

WANG Yun-han1, CHI Ya-nan1, YANG Guan-heng2, FAN Shu-yue1, MA Ji1, XUE Yan1, 2, ZENG Fan-yi1, 2   

  1. 1. Department of Histoembryology, Genetics & Development, Shanghai Jiao Tong University College of Basic Medical Sciences, Shanghai 200025, China; 2. Shanghai Key Laboratory of Embryo and Reproduction Engineering, National Health Commission of the People′s Republic of China Key Laboratory of Medical Embryogenesis and Developmental Molecular Biology, Shanghai Institute of Medical Genetics, Childrens Hospital of Shanghai Jiao Tong University, Shanghai 200040, China
  • Online:2019-11-28 Published:2019-12-16
  • Supported by:
    National Program on Key Basic Research Project of China, 2014CB964701, 2014CB964703; National Natural Science Foundation of China, 31871484; Shanghai Top-Priority Clinical Key Disciplines Construction Project, 2017ZZ02019; Natural Science Foundation of Shanghai, 16ZR1428600

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摘要： 目的·探究不同类型小鼠胎肝基质细胞在细胞形态、分子特征和生物学功能上的差异。方法·通过体外贴壁培养的方式获取13.5 d小鼠胎肝基质细胞，根据细胞形态及其表面标志物CD44、CD29、CD106、CD45及Sca-1的表达差异，区分不同的细胞类型。使用小鼠基因组阵列430 2.0基因芯片在P3或-2的标准下筛选差异基因，并使用IPA（ingenuity pathway analysis）软件对典型信号通路和生物学功能进行分析。结果·依据细胞形态的不同，分离出2类胎肝基质细胞，即短梭形细胞（CD45+CD106-CD29+CD44+Sca-1-）和成纤维样细胞（CD45-CD106+CD29+CD44+Sca-1-）。前者共筛选出1 485个高表达基因，主要参与免疫炎症相关的信号通路；后者共筛选出3 374个高表达基因，主要参与细胞外基质形成及细胞间黏附。结论·小鼠胎肝基质细胞在生物学特征及功能上具有较强的异质性，在促进造血干细胞的增殖方面可能存在差异。

关键词: 胎肝, 基质细胞, 造血干细胞, 流式细胞术, 基因芯片, IPA软件

Abstract:

Objective · To explore the differences in morphology, molecular characteristics and biological functions of different types of mofetal liver stromal cells. Methods · E13.5 mofetal liver stromal cells were obtainedadherent culture, and different cell types were distinguishedmorphology and of surface markers, such as CD44, CD29, CD106, CD45 and Sca-1. Then microarray assay was conductedMoGenome 430 2.0 Array and analyzed at P3 or -2 to identify differential genes. Canonical pathway and biological function analyses were performedusing ingenuity pathway analysis (IPA software). Results · Spindle-like (CD45+CD106-CD29+CD44+Sca-1-) and fibroblast-like (CD45-CD106+CD29+CD44+Sca-1-) fetal liver stromal cells were isolated in this study according to the cell morphology. The 1 485 highly-expressed genes in spindle-like cells mainly involved in immune and inflammation-related signaling pathways; while the 3 374 highly-expressed genes in fibroblast-like cells mainly involved in extracellular matrix formation and cellular adhesion. Conclusion · Mofetal liver stromal cells have strong heterogeneity in biological characteristics and functions, especially in hematopoietic promoting capability.

Key words: fetal liver, stromal cell, hematopoietic stem cell (HSC), flow cytometry, microarray, ingenuity pathway analysis (IPA)

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