半乳糖凝集素在脐带间充质干细胞中的表达

发布时间：2018-04-08 14:18

本文选题：半乳糖凝集素类　切入点：间充质干细胞　出处：《北京大学学报(医学版)》2013年03期

【摘要】：目的:研究人Wharton’s jelly组织来源的脐带间充质干细胞(umbilical cord mesenchymal stem cells,UC-MSCs)中半乳糖凝集素(Galectins)分子的表达情况。方法:无菌条件下取足月剖宫产术后的胎儿脐带,分离Whar-ton’s jelly,组织块法原代培养UC-MSCs。光镜观察获得UC-MSCs形态,用流式细胞术检测其免疫学特性,茜素红钙结节染色及油红O染色鉴定其分化为成骨细胞和脂肪细胞的能力。采用反转录-聚合酶链式反应(reverse tran-scription-polymerase chain reaction,RT-PCR)检测UC-MSCs中Galectins家族分子的mRNA表达情况,酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测UC-MSCs培养上清液中Galectin-3的含量。结果:组织块法原代培养能获得贴壁生长的UC-MSCs,流式细胞术检测提示其高表达CD29/CD44/CD73/CD90及CD105表面标记,低表达造血细胞表面标记CD14、CD34及CD45和免疫排斥相关分子HLA-DR。在特定诱导条件下UC-MSCs具有成骨和成脂分化的能力。UC-MSCs可在mRNA水平表达Galectin-1,3,4,8,9,且能够以细胞数目依赖的方式外分泌Galectin-3,不同密度接种组间差异均有统计学意义(F=16.901,P=0.002),但Galectin-3的分泌不具备时间依赖性。结论:应用组织块法可以从Wharton’jelly组织中成功培养出UC-MSCs,且UC-MSCs可表达分泌型Galectin-3,为进一步研究UC-MSCs的免疫调控机制奠定了基础。
[Abstract]:Aim: to study the expression of galactose lectin (Galectins) in human umbilical cord mesenchymal stem cells (UC-MSCs) derived from human Wharton's jelly tissue.Methods: umbilical cord of fetus after term cesarean section was taken under aseptic condition, Whar-ton's jellywas isolated and UC-MSCs were cultured by tissue block method.The morphology of UC-MSCs was observed under light microscope and its immunological characteristics were detected by flow cytometry. The ability to differentiate into osteoblasts and adipocytes was identified by alizarin red calcium nodule staining and oil red O staining.The mRNA expression of Galectins family in UC-MSCs was detected by reverse tran-scription-polymerase chain reaction reverse polymerase chain reaction (RT-PCR), and the Galectin-3 content in UC-MSCs supernatant was detected by enzyme-linked immunosorbent assay-ELISA-enzyme linked immunosorbent assay (Elisa).Results: UC-MSCs of adherent growth could be obtained in primary culture by tissue mass method. Flow cytometry showed that UC-MSCs expressed high expression of CD29/CD44/CD73/CD90 and CD105, and low expression of CD14, CD34, CD45 and HLA-DR.UC-MSCs have the ability of osteogenesis and adipogenic differentiation under certain inducement conditions. UC-MSCs can express Galectin-1P0. #number0# at the mRNA level, and excrete Galectin-3 in a manner dependent on the number of cells. There are significant differences among different density inoculation groups, but the secretion of Galectin-3 is significant.No time dependence.Conclusion: UC-MSCs can be successfully cultured from Wharton'jelly tissue by tissue block method, and UC-MSCs can express secretory Galectin-3, which lays a foundation for further study of immunological regulation mechanism of UC-MSCs.
【作者单位】：北京大学第三医院风湿免疫科;
【基金】：国家自然科学基金(81072474,81273293) 北京市自然科学基金(7112143)资助~~
【分类号】：R329

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