中国组织工程研究2024，Vol.28Issue(31)：4926-4930,5.DOI:10.12307/2024.702
长期传代培养对骨髓间充质干细胞生物学特性的影响
Effects of long-term subculture on biological characteristics of bone marrow mesenchymal stem cells
摘要
Abstract
BACKGROUND:There is still controversy whether human bone marrow mesenchymal stem cells can maintain their biological characteristics,energy metabolism patterns,and multidirectional differentiation potential after long-term expression in vitro.Further comprehensive and systematic research is needed. OBJECTIVE:To evaluate the effects of long-term expansion in vitro on the biological characteristics of mesenchymal stem cells. METHODS:Human bone marrow mesenchymal stem cells cultured to passage 5,10,and 15 in vitro.MTT assay was used to detect cell proliferation ability.Flow cytometry was used to detect cell cycle.The multi-differentiation potential of mesenchymal stem cells was detected by inducing to adipogenic,osteogenic and chondrogenic differentiation.Cell migration and invasion abilities were detected by scratch test and Transwell assay.The mitochondrial oxidative phosphorylation and glycolysis function were analyzed using energy metabolism analyzer.The cell senescence was detected by senescence-associated β-galactosidase staining.The expression levels of p21,p16,and p53 proteins were detected by western blot assay. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells at passages 5,10,and 15 grew adherently;the volume of passage 15 mesenchymal stem cells increased and its proliferation ability decreased;the percentage of S-phase cells decreased(P<0.05).With the increase of culture passages,the migration and invasion abilities decreased gradually(P<0.05).There was no significant difference in the differentiation potential,demonstrated by adipogenic,osteogenic and chondrogenesis induction.The ability of oxidative phosphorylation of mitochondria and glycolysis decreased gradually(P<0.05).The number of senescence-associated β-galactosidase-positive cells increased with the increase of passages(P<0.05),and the expression of senescence protein p21,p16,and p53 increased gradually(P<0.05).The results indicated that the biological characterization of mesenchymal stem cells changed after long-term in vitro expansion.Mesenchymal stem cells cultured over 10 passages may have a reduced activity due to increasing senescence.Therefore,bone marrow mesenchymal stem cells cultured less than 10 passages are suitable for clinical research/therapy.关键词
骨髓间充质干细胞/传代/长期培养/能量代谢/衰老Key words
bone marrow mesenchymal stem cell/passage/long-term culture/energy metabolism/senescence分类
医药卫生引用本文复制引用
赵文静,刘百坤,李秋莲,陈曦..长期传代培养对骨髓间充质干细胞生物学特性的影响[J].中国组织工程研究,2024,28(31):4926-4930,5.基金项目
吉林省卫生健康科技能力提升项目(2020J082),项目参与人:赵文静 （2020J082）
吉林省卫生健康科技能力提升项目(2022JC072),项目参与人:赵文静Jilin Province Health Technology Capability Enhancement Project,No.2020J082(to ZWJ) （2022JC072）
Jilin Province Health Technology Capability Enhancement Project,No.2022JC072(to ZWJ) （to ZWJ）
