首页|小鼠骨髓和脂肪间充质干细胞定向分化能力的比较研究

小鼠骨髓和脂肪间充质干细胞定向分化能力的比较研究

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目的 探讨小鼠骨髓源性间充质干细胞(BM-MSCs)和脂肪源性间充质干细胞(AD-MSCs)的定向分化能力.方法 从C57BL/6J小鼠股骨骨髓和腹股沟白色脂肪组织中分别分离和培养BM-MSCs和AD-MSCs,分别使用成骨、成软骨和成脂诱导分化培养基诱导两种细胞定向分化.采用茜素红、阿利新蓝和油红O染色检测成骨、成软骨和成脂分化程度;实时荧光定量PCR(qPCR)鉴定MSCs并检测定向分化相关基因Runx2、Sp7(成骨),Sox9、Col2a1(成软骨),Pparg和Cebpa(成脂)表达水平,确定细胞的定向分化能力.基于GEO数据库中GSE43804和GSE122778数据集的小鼠和人类BM-MSCs和AD-MSCs基因表达谱数据,分析差异表达基因及其富集的信号通路.结果 分离培养得到的BM-MSCs和AD-MSCs细胞形态不同,AD-MSCs梭形形态更明显;两种细胞均表达CD29、CD44和CD90,不表达CD34和CD45.定向诱导后AD-MSCs的成骨和成脂分化程度高于BM-MSCs,而成软骨分化程度低于BM-MSCs(P<0.05);定向诱导后AD-MSCs中Runx2、Pparg和Cebpa mRNA表达水平高于BM-MSCs,Sox9 mRNA表达水平低于BM-MSCs(P<0.05).小鼠和人的AD-MSCs高表达的基因富集于PPAR和WNT信号通路,BM-MSCs高表达的基因富集于软骨和骨发育信号通路.结论 小鼠AD-MSCs成骨和成脂分化能力强于BM-MSCs,而成软骨分化能力弱于BM-MSCs,PPAR、WNT、软骨和骨发育信号通路的活化状态在决定BM-MSCs和AD-MSCs不同定向分化潜能中起重要调节作用.
Comparative study on the directed differentiation ability of mouse bone marrow and adipose-derived mesenchymal stem cells
Objective To investigate the targeted differentiation ability of mouse bone marrow derived mesenchymal stem cells(BM-MSCs)and adipose-derived mesenchymal stem cells(AD-MSCs).Methods BM-MSCs and AD-MSCs were isolated and cultured from bone marrow of femur and white adipose tissue of groin of C57BL/6J mice respectively,and the two types of cells were induced by osteogenic,chondrogenic and adipogenic differentiation medium respectively.Alizarin red,alcian blue and oil red O staining were used to detect the differentiated degree of osteogenic,chondrogenic and lipogenic differentiation.Real-time fluorescence quantitative PCR(qPCR)was used to identify MSCs and detected expression levels of directed differentiation-related genes Runx2,Sp7(osteoblast),Sox9,Col2a1(chondroblast),Pparg and Cebpa(lipogenesis)to determine the directed differentiation ability of cells.Based on gene expression profiles of mouse and human BM-MSCs and AD-MSCs in GEO database GSE43804 and GSE122778,the differentially expressed genes and their enrichment signal pathways were analyzed.Results The cell morphology of BM-MSCs and AD-MSCs obtained by isolation and culture was different,and spindle-shaped morphology was more obvious in AD-MSCs.Both cells expressed CD29,CD44 and CD90,but did not express CD34 and CD45.AD-MSCs showed higher osteogenic and lipogenic differentiation than those of BM-MSCs after directed induction,while chondrogenic differentiation was lower in AD-MSCs than that of BM-MSCs(P<0.05).After directional induction,expression levels of Runx2,Pparg and Cebpa mRNA were higher in AD-MSCs than those in BM-MSCs,and Sox9 mRNA expression levels were lower than those in BM-MSCs(P<0.05).Highly expressed genes of AD-MSCs in mice and human were enriched in PPAR and WNT signaling pathways.Highly expressed genes of BM-MSCs were enriched in cartilage and bone developmental signaling pathways.Conclusion The osteogenic and adipogenic differentiation ability of mouse AD-MSCs is stronger than those of BM-MSCs,while the chondrogenic differentiation ability AD-MSCs is weaker than that of BM-MSCs.The activation status of PPAR,WNT,cartilages and skeletal system development signaling pathways plays an important regulatory role in determining the different directional differentiation potential of AD-MSCs and BM-MSCs.

mesenchymal stem cellsbone marrowfatsPPAR gammaWnt signaling pathwayosteogenic differentiationchondrogenic differentiationadipogenic differentiation

钟家帅、冯玉梅

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天津医科大学肿瘤医院,国家恶性肿瘤临床医学研究中心,天津市恶性肿瘤临床医学研究中心,天津市肿瘤防治重点实验室,天津医科大学肿瘤医院肿瘤研究所生物化学与分子生物学研究室(邮编 300060)

间质干细胞 骨髓 脂肪类 PPARγ Wnt信号通路 成骨分化 成软骨分化 成脂分化

天津市医学重点学科(专科)建设项目

TJYXZDXK-009A

2024

天津医药
天津市医学科学技术信息研究所

天津医药

CSTPCD
影响因子:1.107
ISSN:0253-9896
年,卷(期):2024.52(2)
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