滩羊原代前体脂肪细胞不同培养方法的比较OA

To screen an efficient and stable ovine preadipocyte culture method,tail adipose tissue from 6-month-old Tan sheep was used as the material in this study.EdU,CCK-8,Nile Red,and Oil Red O were used to comprehensively compare the proliferation ability,viability,and adipogenic differenti-ation ability of cells at different passages using three investigated methods(enzyme digestion,tissue granule,and tissue block).The expression of adipogenic marker genes,PPARγ andFABP4 at differ-ent passages was assessed by RT-qPCR.The results showed that the three methods yielded cells with typical spindleor star-shaped morphology and high purity(positive for PPARγ immunofluorescence).The tissue granule method had significantly higher separation efficiency than the tissue block method(i.e.,shorter time required)but was less efficient than enzymatic digestion.At early passages(P3),cell proliferation was significantly higher in the enzymatic digestion and tissue block groups than in the tissue granule group(P＜0.05),and adipogenic potential was significantly greater in the tissue block group than in the tissue granule group(P＜0.01);at later passages(P9),the tissue block groups showed significantly higher proliferation ability than the enzymatic digestion group(P＜0.01),while the tissue block group exhibited the lowest adipogenic capacity.With increasing passage number,both proliferation and adipogenic abilities declined across all three methods.However,the tissue granule method exhibited the slowest decline in proliferation(P＜0.05)and maintained strong adipogenic ca-pacity at P7 and P9(P＜0.01).In conclusion,the tissue granule method shows the best performance in maintaining the proliferation stability and adipogenic potential of later-passage cells.