脐带间充质干细胞体外衰老过程中生物学特性及基因表达谱的变化  

Changes of Biological Characteristics and Gene Expression Profile of Umbilical Cord Mesenchymal Stem Cells During Senescence in Culture

作　　者：宁雪[1] 李栋[1] 汪大琨[1] 付金秋[1] 鞠秀丽[1] 

机构地区：[1]山东大学齐鲁医院儿童医疗中心,山东济南250012

出　　处：《中国实验血液学杂志》2012年第2期458-465,共8页Journal of Experimental Hematology

基　　金：山东省科技发展计划项目;编号2009GG10002027

摘　　要：本研究探讨人脐带间充质干细胞(UC-MSC)在体外培养的衰老过程中细胞生物学特征及衰老相关基因的变化特点。体外分离培养UC-MSC,取第3代(对照组)和第15代(衰老组)细胞进行形态学观察、增殖检测、流式表型测定和人类全基因组表达谱芯片分析,并选取重要基因进行定量逆转录多聚酶链反应验证。结果显示,与对照组相比,衰老组细胞形态变大,增殖速度减慢,但CD44和CD105等细胞表型阳性率无变化;衰老细胞组的核糖体小亚基组成相关基因显著上调,上调的信号通路主要涉及类固醇合成、半乳糖代谢、自身免疫病和退行性疾病;下调明显的是细胞骨架,DNA、mRNA结构的结合以及蛋白质功能等相关基因,与细胞黏附功能和细胞增殖周期等相关。结论:第15代的UC-MSC出现细胞代谢与增殖功能下降,从而导致细胞衰老。This study was purposed to investigate the changes of biological properties and expression patterns of the aging related genes in umbilical cord mesenchymal stem cells （UC-MSC） during in vitro culture. UC-MSC at passage 3 were served as the control cells and those at passage 15 were considered as the aged cells. The biological features of those two kinds of cells including morphology, proliferation activity and phenotypic profile were observed, and the differences of gene expression were analysed by the whole human genome oligo microarray. Several differential genes were selected for further confirmation by quantitative reverse transcription-polymerase chain reaction. The results showed that UC-MSC at passage 15 were larger in size and their proliferation rate was slower compared with those of cells at passage 3, while the positivity of CD44 and CD105 remained unchanged. Compared with UC-MSC at passage 3, relatively aged cells expressed higher levels of genes that are associated with small subunit of ribosome. Further analysis with Gene Ontology functional categories showed that the up-regulated genes were concentrated in those related to steroid biosynthesis, galactose metabolism and the development of autoimmune diseases and degenerative diseases and the down-regulated genes in UC-MSC at passage 15 were concentrated in cytoskeleton molecules, DNA structure binding, mRNA binding and protein function. Functional analysis with Kyoto Encyclopedia of Genes and Genomes functional pathway revealed that the expression of some genes responsible for ribosome composition was elevated while those of associated with extracellular matrix,focal adhesion and cell cycle progression were down-regulated. It is concluded that UC-MSC become senescent due to the declines in metabolism and proliferation activities.

关 键 词：脐带间充质干细胞 衰老 基因表达 信号通路 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]