BMSC细胞外基质对脂肪间充质干细胞生物学活性的影响  

Effect of BMSC derived extracellular matrix on biological activity of adipose mesenchymal stem cells

作　　者：原芳芳 王丽[1] 刘伟[2] 吴松芳 郭红延 YUAN Fang-fang;WANG Li;LIU Wei;WU Song-fang;GUO Hong-yan(Department of Stomatology,The Third Medical Center of Chinese PLA General Hosptial,Beijing 100039,China)

机构地区：[1]解放军总医院第三医学中心口腔科,北京100039 [2]军事科学院军事医学研究所,北京100850

出　　处：《中华老年口腔医学杂志》2020年第2期97-102,共6页Chinese Journal of Geriatric Dentistry

基　　金：国家自然青年科学基金(项目编号:31700843)。

摘　　要：目的:探讨骨髓间充质干细胞(BMSCs)衍生的细胞外基质(Extracellular matrix,ECM)对脂肪间充质干细胞(Adipose stem cells,ADSCs)增殖及多向分化能力的影响。方法:制备BMSCs衍生的ECM材料(BM-dECM),利用H&E染色、免疫荧光染色以及DNA检测方法对BM-dECM进行表征。将ADSCs分别接种于BM-dECM包被的培养皿以及空白培养皿中,利用细胞计数试剂盒(CCK-8)检测两组ADSCs的增殖能力;利用RT-PCR检测两组ADSCs成骨、成脂以及成软骨分化相关基因的表达。结果:H&E染色结果显示BM-dECM呈连续的膜状结构,细胞核被大部分脱除;免疫荧光染色显示BM-dECM保留了BMSCs分泌的细胞外基质成分;DNA含量检测证明脱细胞后ECM中DNA含量下降为17.72±3.24ng/μl;CCK-8检测显示,与对照组相比BM-dECM显著促进ADSCs增殖;RT-PCR结果显示BM-dECM在一定程度上促进了ADSCs成骨分化相关基因的表达,抑制了成脂以及成软骨相关基因的表达。结论:BM-dECM可以有效促进ADSCs增殖,抑制ADSCs成脂以及成软骨能力而增强其成骨分化的能力。Objective:To investigate the effects of extracellular matrix(ECM)derived from bone marrow mesenchymal stem cells(BMSCs)on the proliferation and multi-directional differentiation of adipose mesenchymal stem cells(ADSCs).Methods:BMSCs-derived ECM material(BM-d ECM)was prepared and characterized by hematoxylin-eosin staining,immunofluorescence staining and Genomic DNA Kit.ADSCs were cultured into BM-dECM-coated culture dishes and blank culture dishes respectively,and the proliferation of the ADSCs was detected by cell counting kit-8(CCK-8)was used to detect the expression of adipogenic and chondrogenic differentiation related genes in two groups of ADSCs.Results:H&E staining showed that BM-dECM had a continuous membrane structure with most of nucleus removed;Immunofluorescence staining showed that BM-dECM retained the extracellular matrix component secreted by BMSCs;DNA content testing showed that the amount of DNA in ECM decreased to 17.72±3.24 ng/μl after decellularization;CCK-8 assay showed that BM-dECM significantly promoted the proliferation of ADSCs compared with the control group(P<0.05);RT-PCR results showed that BM-d ECM promoted the expression osteogenic differentiation related genes of ADSCs,and inhibited the expression of adipogenic and chondrogenic related genes.Conclusion:BM-dECM can effectively promote the proliferation of ADSCs and enhance the potential of osteogenic differentiation.

关 键 词：细胞外基质 骨髓间充质干细胞 脂肪间充质干细胞 细胞增殖 细胞分化 

分 类 号：R781.4[医药卫生—口腔医学]