Marc-145细胞的无血清悬浮培养基开发

Development of serum-free suspension medium for Marc-145 cells

为开发适合Marc-145细胞生长的无血清悬浮培养基,试验将贴壁Marc-145细胞转入Celer-S001培养基中进行悬浮适应,通过混料试验和水解物筛选试验获得适应Marc-145细胞快速扩增的无血清悬浮培养基,对悬浮培养的Marc-145细胞进行猪繁殖与呼吸综合征病毒感染并检测病毒效含量,评价悬浮培养的Marc-145细胞的病毒扩增能力。结果表明:Marc-145细胞可以在Celer-S001培养基中悬浮培养,其比生长速率为(0.25±0.06)/d;适应Marc-145细胞快速扩增的最优无血清悬浮培养基由B3和B8培养基以0.54∶0.46比例混合,并添加2 g/L的水解物H3构成,Marc-145细胞的比生长速率为(0.51±0.03)/d,密度为3.73×10^(6) cells/mL;细胞接毒后的病毒含量最高可达(5.25±0.25)lgTCID_(50)/mL,表明细胞仍然保持着病毒扩增能力。说明试验成功开发了适合Marc-145细胞生长的无血清悬浮培养基,Marc-145细胞在该培养基中生长良好,具有病毒扩增能力。

In order to develop serum-free suspension medium for Marc-145 cells,in this experiment adherent Marc-145 cells were transferred into Celer-S001 medium for suspension adaptation.A serum-free suspension medium suitable for rapid proliferation of Marc-145 cells was obtained by mixture test and hydrolysate screening test.Suspension cultured Marc-145 cells were infected with Porcine reproductive and respiratory syndrome virus and the virus titer was detected to evaluate the virus proliferation ability of suspension cultured Marc-145 cells.The results showed that Marc-145 cells could be cultured in suspension in Celer-S001 medium with a specific growth rate of(0.25±0.06)/d.The optimal serum-free suspension medium for the rapid proliferation of Marc-145 cells was composed of B3 and B8 medium mixed in a ratio of 0.54∶0.46 and supplemented with 2 g/L of hydrolyzate H3;the specific growth rate of Marc-145 cells was(0.51±0.03)/d,and the density of Marc-145 cells was 3.73×10^(6) cells/mL.The virus titer after inoculation of the cells was up to(5.25±0.25)lgTCID_(50)/mL,indicating that the cells still maintained the ability of virus proliferation.The results suggested that a serum-free suspension medium suitable for the growth of Marc-145 cells had been successfully developed.Marc-145 cells growed well in this medium,and the cultured suspension Marc-145 cells had virus proliferation ability.