2类小鼠胎肝基质细胞的生物学特征及表达谱分析  

Biological characteristics and gene expression profile analysis of two kinds of mouse fetal liver stromal cells

作　　者：王沄汉 迟亚男 杨冠恒[2] 范书玥 马姬 薛燕 曾凡一[1,2] WANG Yun-han;CHI Ya-nan;YANG Guan-heng;FAN Shu-yue;MA Ji;XUE Yan;ZENG Fan-yi(Department of Histoembryology,Genetics&Development,Shanghai Jiao Tong University College of Basic Medical Sciences,Shanghai 200025,China;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,Children's Hospital of Shanghai Jiao Tong University,Shanghai 200040,China)

机构地区：[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　　处：《上海交通大学学报（医学版）》2019年第11期1226-1232,共7页Journal of Shanghai Jiao tong University:Medical Science

基　　金：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)~~

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4302.0基因芯片在P<0.05、差异倍数>3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1485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3374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。Objective·To explore the differences in morphology,molecular characteristics and biological functions of different types of mouse fetal liver stromal cells.Methods·E13.5 mouse fetal liver stromal cells were obtained by adherent culture,and different cell types were distinguished by morphology and expression of surface markers,such as CD44,CD29,CD106,CD45 and Sca-1.Then microarray assay was conducted by Mouse Genome 4302.0 Array and analyzed at P<0.05,fold change>3 or<-2 to identify differential expression genes.Canonical pathway and biological function analyses were performed by using 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 1485 highly-expressed genes in spindle-like cells mainly involved in immune and inflammation-related signaling pathways;while the 3374 highly-expressed genes in fibroblastlike cells mainly involved in extracellular matrix formation and cellular adhesion.Conclusion·Mouse fetal liver stromal cells have strong heterogeneity in biological characteristics and functions,especially in hematopoietic promoting capability.

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 

分 类 号：Q291[生物学—细胞生物学]