基于细胞体外培养筛选最适胎牛血清

Screening the Optimal Fetal Bovine Serum Based on in vitro Cell Culture

为了探索小鼠骨髓瘤细胞(Sp2/0)与猪胎儿成纤维细胞(PFF)对胎牛血清(FBS)的需求差异,以期筛选出最适血清,首先采用两种不同来源的FBS培养Sp2/0和PFF,观察细胞形态、克隆形成,分析细胞活率与数量.结果显示:无论采用国产FBS1还是进口的FBS2培养Sp2/0,其细胞克隆数、细胞数量和活率差异均无统计学意义(p>0.05),但进口FBS2培养的PFF,其细胞克隆数、细胞数量和活率均显著高于国产FBS1(p<0.05).随后用进口FBS2的3个不同批号培养PFF,结果显示:3个批号的FBS2均能促进细胞生长,细胞结构清晰,呈梭形;培养7 d,3个批次的FBS2均有细胞克隆形成,但FBS2-1形成的克隆小,且细胞克隆数极显著低于FBS2-2和FBS2-3(p<0.01);培养10 d,FBS2-1的细胞数量和细胞活率极显著低于FBS2-2和FBS2-3(p<0.01).虽然FBS2-2的细胞克隆数极显著低于FBS2-3(p<0.01),但是FBS2-2的细胞克隆大,且细胞数量极显著高于FBS2-3(p<0.01).结果表明:Sp2/0与PFF对FBS的需求有差异,无论是国产FBS1还是进口FBS2,均能满足Sp2/0体外培养的需要,但PFF在进口FBS2中的生长性能极显著优于国产FBS1,FBS2-2是PFF体外培养的最适血清.

In order to explore the difference in the demand for fetal bovine serum(FBS)between mouse myeloma cells(Sp2/0)and porcine fetal fibroblasts(PFF),and screen the optimal serum for the two cells,in this study,Sp2/0 and PFF cells were cultured with two different sources of FBS.The cell viability and number were analyzed by observing the cell morphology and clone formation.The results showed that there was no significant difference in the number of cell clone,the number of cell and cell viability between domestic FBS1 and imported FBS2(p>0.05).However,the number of cell clone,the number of cell and cell viability of PFF,which cultured with FBS2 were significantly higher than those of domestic FBS1(p<0.05).Subsequently,PFF was cultured with three different batches of imported FBS2.The results showed that FBS2 of three batches could promote cell growth,and the cell structure was clear and spindle shaped.After 7 days of culture,cell clones were formed in three batches of FBS2,but the clones formed in FBS2-1 were small,and the number of cell clones was significantly lower than that of FBS2-2 and FBS2-3(p<0.01).After 10 days of culture,the number of cell and cell viability of FBS2-1 were significantly lower than those of FBS2-2 and FBS2-3(p<0.01).Although the number of cell clones of FBS2-2 was significantly lower than that of FBS2-3(p<0.01),but the number of cell of FBS2-2 was significantly higher than that of FBS2-3(p<0.01).The study reveals that there are differences in the demand for FBS between Sp2/0 and PFF.Both domestic FBS1 and imported FBS2 can meet the needs of Sp2/0 in vitro culture,but the growth performance of PFF in imported FBS2 is significantly better than that of domestic FBS1,and FBS2-2 is the optimal serum for PFF in vitro culture.