The development of virus-free, oral vaccines against poliovirus capable of inducing mucosal protective immunity is needed to safely combat this pathogen. In the present study, a carrot cell line expressing the poliovirus VP2 antigen was established at the level of callus and cell suspensions, exploring the effects of culture media (MS and B5), supplementation with urea, phytoregulators (2,4-D : KIN), and light conditions (continuous light, photoperiod, and total darkness). The best callus growth was obtained on B5 medium supplemented with 2 mg/L of 2,4-D + 2 mg/L kinetin and 0.0136 g/L of urea and in continuous light conditions. Suspension cultures of the SMC-1 line in 250 mL Erlenmeyer flasks had a maximum growth of 16.07 ± 0.03 g/L DW on day 12 with a growth rate of µ=0.3/d and a doubling time of 2.3 days. In a 2 L airlift bioreactor, the biomass yield achieved was 25.6 ± 0.05 g/L DW at day 10 with a growth rate of µ= 0.58/d and doubling time of 1.38 d. Cell growth was 1.5 times higher in bioreactors than in shake flasks, highlighting that both systems resulted in the accumulation of VP2 throughout the time in culture. The maximum VP2 yield in flasks was 387.8 µg/g DW at day 21, while in the reactor it was 550.2 µg/g DW at day 18. In conclusion, bioreactor-based production of the VP2 protein by the SMC-1 suspension cell line offers a higher productivity when compared to flask cultures, offering a key perspective to produce low-cost vaccines against poliomyelitis.

为了安全地对抗这种病原体，需要开发能够诱导粘膜保护性免疫的无病毒口服脊髓灰质炎病毒疫苗。在本研究中，在愈伤组织和细胞悬浮液水平上建立了表达脊髓灰质炎病毒VP2抗原的胡萝卜细胞系，探索了培养基（MS和B5）、补充尿素、植物调节剂（2,4-D：KIN ）和光照条件（连续光照、光周期和完全黑暗）。在补充有 2 mg/L 2,4-D + 2 mg/L 激动素和 0.0136 g/L 尿素的 B5 培养基上并在连续光照条件下获得最佳愈伤组织生长。SMC-1 系在 250 mL 锥形烧瓶中的悬浮培养物在第 12 天的最大生长量为 16.07 ± 0.03 g/L DW，生长速率为 µ=0.3/d，倍增时间为 2.3 天。在 2 L 气升式生物反应器中，第 10 天实现的生物量产量为 25.6 ± 0.05 g/L DW，增长率为 µ= 0.58/d，倍增时间为 1.38 d。生物反应器中的细胞生长速度比摇瓶中的细胞生长速度高 1.5 倍，这突出表明这两种系统都会导致 VP2 在培养过程中不断积累。第 21 天，烧瓶中的最大 VP2 产量为 387.8 µg/g DW，而在反应器中，第 18 天的最大 VP2 产量为 550.2 µg/g DW。 总之，SMC-1 悬浮细胞系基于生物反应器生产 VP2 蛋白与烧瓶培养相比，它具有更高的生产率，为生产低成本的脊髓灰质炎疫苗提供了关键视角。