HepG2细胞培养方法与条件的探讨

The Discussion on HepG2 Cell Culture Method and Condition

目的 探讨HepG2细胞最优培养方法与培养条件。 方法 对比 40℃先溶解后 3 7℃水浴恒温与传统 3 7℃复苏方法效果以选择最佳的复苏方法 ;观察不同比例胎牛血清培养基下细胞生长情况以确定最佳的血清含量 ;观察不同汇合度下细胞生长趋势及死活细胞比例以选择恰当的传代时机 ;观察不同消化方法的消化效果以选择合适的消化方法。 结果 40℃先溶解后 3 7℃水浴恒温的复苏存活率 (85 .7% )明显高于传统 3 7℃复苏方法 (68.4% ) ;在含 12 %以上胎牛血清的DMEM中细胞生长良好 ;汇合度达 95 %～ 10 0 %时细胞生长趋势达到平台期 ,10 0 %以后活细胞逐渐减少而死细胞逐渐增加 ;应用先PBS洗涤后 0 .2 5 %胰酶消化的传代方法易于控制 ,消化效果好。 结论 HepG2细胞采取 40℃先溶解后 3 7℃水浴恒温的复苏方法 ,含 12 %胎牛血清DMEM培养基培养 ,采用先PBS洗涤后胰酶消化的方法于汇合度 95 %～ 10 0 %时传代的方法进行培养 ,能获得最佳培养效果。

Objective To probe into the best culture technique and condition of HepG2. Methods Compare the resuscitation effect of dissolving in 40 ℃ advance plus 37 ℃ with the tradition 37 ℃ resuscitation method. Confirm the best serum content by observing the grow circus of HepG2 in different serum content culture medium. Choose the best separate time by observing the cell grown trend and proportion of life and death cell in different convergence degree. Choose the appropriate digest method by observing the digest effect of different digest method. Results The resuscitation livability of dissolve in 40 ℃ advance plus 37 ℃ resuscitation method (85.7%) is evidently higher than the tradition 37 ℃ resuscitation method( 68.4%). Cells grow well in DMEM that contain 10% fetal cattle serum. The trend of growth reach flat when convergence degree is 95%～100% . Life cells drop off and death cells gradually increase when convergence degree reached 100%. It is easy to control and have good digest effect that use PBS wash the cells advance then apply 0.25%trypsin to digest. Conclusion The culture of HepG2 can apply the 40 ℃ advance plus 37 ℃ resuscitation method, growing in DMEM that contain 12% fetal cattle serum, and apply PBS wash advance then 0.25% trypsin to digest when convergence degree reachs 95%～100%.