长白猪骨髓和脂肪间充质干细胞复制性衰老过程中生物学特性和抗氧化损伤能力的比较  

Comparative Study of Biological Characteristics and Anti Oxidative Damage Ability of Porcine Bone Marrow and Adipose Derived Mesenchymal Stem Cells During the Replicative Senescence

作　　者：车丽娜 李凤娇 赵崇杰 黄磊[1] 许慧 王楠[1] CHE Lina;LI Fengjiao;ZHAO Chongjie;HUANG Lei;XU Hui;WANG Nan(College of Biotechnology,Tianjin University of Science&Technology,Tianjin 300457,China)

机构地区：[1]天津科技大学生物工程学院,天津300457

出　　处：《天津科技大学学报》2023年第2期1-10,共10页Journal of Tianjin University of Science & Technology

基　　金：国家重点研发计划项目(2021YFC2101403)。

摘　　要：为研究对比长白猪骨髓间充质干细胞(bone marrow derived mesenchymal stem cells,BMSCs)和脂肪间充质干细胞(adiposederivedmesenchymalstemcells,AMSCs)在体外衰老过程中的生物学特性和抗氧化损伤能力,分别利用贴壁筛选法、Ⅰ型胶原酶消化法分离获得BMSCs和AMSCs,利用流式细胞术鉴定细胞,EdU检测细胞增殖能力,实时荧光定量聚合酶链反应(RT-qPCR)检测基因表达,β-半乳糖苷酶(β-galactosidase,β-gal)染色检测细胞衰老情况,油红O和茜素红染色检测细胞的成脂和成骨分化能力,TUNEL检测细胞的凋亡情况.分离的BMSCs和AMSCs均高表达CD44和CD90,不表达CD45和CD34.与AMSCs相比,P8代BMSCs增殖能力强,OCT4和Ki67表达高,β-gal阳性细胞数少,成骨分化能力高.但在成脂分化能力方面,AMSCs强于BMSCs.进一步发现H_(2)O_(2)处理P8代细胞时,AMSCs比BMSCs对氧化损伤更敏感,细胞凋亡率显著增加,究其原因可能与抗氧化损伤基因表达的显著降低有关.In order to study and compare the biological characteristics and antioxidant capacity of bone marrow derived mesenchymal stem cells(BMSCs)and adipose derived mesenchymal stem cells(AMSCs)from Landrace pigs during aging in vitro,BMSCs and AMSCs were isolated by adherent screening method and typeⅠcollagenase digestion method respectively.MSCs were identified by using flow cytometry and it’s proliferation capacity was detected with EdU.The expression of gene was detected by RT-qPCR,cell senescence was detected by β-galactosidase(β-gal)staining,the abilities of adipogenic and osteogenic differentiation of these cells were identified by oil red O staining and alizarin red staining,respectively.The apoptosis of cells was detected by TUNEL method.Both the isolated BMSCs and AMSCs highly expressed CD44 and CD90,but not CD45 and CD34.Compared with AMSCs,BMSCs at P8 had strong proliferative ability,increased expression of OCT4 and Ki67,lower numbers of β-gal positive cells,and higher osteogenic differentiation ability.However,adipogenic differentiation ability of AMSCs were stronger than that of BMSCs.Moreover,when P8 cells were treated with H_(2)O_(2),AMSCs were more sensitive to oxidative damage than BMSCs,and the apoptosis rate increased significantly.This may be due to the significant reduction of antioxidant gene expression.

关 键 词：骨髓间充质干细胞 脂肪间充质干细胞 增殖 衰老 抗氧化损伤 生物学特性 

分 类 号：S828.91[农业科学—畜牧学]