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目的探讨应用96孔板悬浮法诱导P19细胞体外向脂肪细胞分化的可行性和有效性。方法 P19细胞使用96孔板悬浮培养法制备拟胚体(EBs),其中第3天至第5天使用诱导剂全反式维甲酸(RA)。第7天挑取EBs用胰岛素和三碘甲状腺原氨酸诱导,继续贴壁培养20d后,油红O染色鉴定分化细胞的特征。结果使用96孔板悬浮培养法可以形成大小均一的EBs,诱导结束后EBs生长晕周围部分细胞分化为脂肪细胞。细胞形态趋于类圆或圆形,胞质中出现小脂滴,可被油红O染色清晰显示。结论使用96孔板悬浮法获得的P19细胞EBs在体外可诱导分化为脂肪细胞。
Objective To investigate the feasibility and effectiveness of using 96-well plate suspension to induce differentiation of P19 cells into adipocytes. Methods P19 cells The embryoid bodies (EBs) were prepared by suspension culture in 96-well plates. The inducer All-trans retinoic acid (RA) was used from day 3 to day 5. On the 7th day, EBs were induced by insulin and triiodothyronine. After adherent culture for 20 days, the characteristics of differentiated cells were identified by oil red O staining. Results The uniform size of EBs could be formed by using 96-well plate suspension culture method. After the induction, EBs grew to differentiate into adipocytes. Cell morphology tends to be round or circular, small lipid droplets appear in the cytoplasm, can be clearly displayed by oil red O staining. Conclusion EBs derived from P19 cells obtained by 96-well plate suspension can be induced to differentiate into adipocytes in vitro.