青年和老年小鼠棕色脂肪来源间充质干细胞生物学特性比较

Comparison of Biological Characteristics of Brown Adipose-derived Mesenchymal Stem Cells in Young and Elder Mice

旨在探讨青年和老年小鼠肩胛骨间棕色脂肪来源间充质干细胞(Brown adipose-derived mesenchymal stem cells,BADSCs)生物学特性的比较。用细胞流式检测第3代BADSCs表面抗原表达,MTT法检测细胞增殖情况,7-ADD-AnnexinV双染色观察细胞凋亡情况和β-半乳糖苷酶染色法检测细胞衰老情况,比较两组细胞的成骨和成脂分化差异。结果显示,第3代BADSCs表面抗原CD29、CD105和CD44的表达均为阳性、CD73为低表达。MTT法显示老年小鼠的BADSCs增殖能力与青年小鼠相比没有显著性差异(P>0.05)。7-ADD-AnnexinV双染色显示青年小鼠的BADSCs早凋比例4.63±0.87%,老年的BADSCs早凋比例9.88±0.81%,且有显著性差异(P<0.05)。β-半乳糖苷酶染色法显示青年组染成蓝色细胞数为2.33±0.3,老年组染成蓝色细胞数为6.66±1.2,有显著性差异(P<0.05)。青年组成骨分化OD值为1.26±0.046,老年组成骨分化OD值为0.88±0.047,有显著性差异(P<0.05)。青年组成脂分化OD值为0.9808±0.066,老年组成脂分化OD值为0.769±0.035,有显著性差异(P<0.05)。青年组的BADSCs更倾向于成骨分化,老年组的BADSCs更倾向于成脂分化。老年组小鼠凋亡和衰老细胞比例增加;成骨和成脂分化能力下降。

This work aims to compare the biological characteristics of brown adipose-derived mesenchymal stem cells(BADSCs)between young and elder mice.Cell flow was used to detect the third-generation BADSCs surface antigen expression,MTT test to determine cell proliferation,7-ADD-AnnexinV double staining to observe the apoptosis,β-glucoside enzyme staining method to detect BADSCs cell aging,and osteogenesis and adiposesis differentiation between 2 groups were compared.As results,the expression levels of surface antigen CD29,CD105 and CD44 of the 3rd generation BADSCs in the young and elder mice were all positive,while CD73 was in low expression.The MTT method showed that the proliferation capacity of BADSCs in the elder mice was not significantly different from that in the young ones.7-ADDAnnexinV double staining demonstrated that the proportion of BADSCs early apoptosis in the young mice was 4.63±0.87%,and that for the elder ones was 9.88±0.81%,i.e.,there was significant difference(P<0.05).The dyed blue cell number in the young group was 2.33±0.3,and that in the elder group was 6.66±1.2,i.e.,difference was significant(P<0.05).The OD value of osteogenetic differentiation in the young group was 1.26±0.066,and that in the elder group was 0.88±0.047,thus this was significantly different(P<0.05).The OD value of adipogenic differentiation in the young group was 0.9808±0.066,and that in the elder group was 0.769±0.035,i.e.,difference was significant(P<0.05).In conclusion,BADSCs in the young group were more inclined to osteogenetic differentiation,while BADSCs in the elder group was more inclined to adipogenic differentiation;the proportion of apoptosis and senescence cells in the elder mice increased,while osteogenesis and adipogenesis decreased.