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Planta Med 2024; 90(01): 63-72
DOI: 10.1055/a-2181-2886
Natural Product Chemistry and Analytical Studies
Original Papers

Establishment of the Daucus carota SMC-1 Cell Suspension Line for Poliovirus Vaccine Development

Mónica Morales-Aguilar
1   Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, Mexico
,
Omayra Citlalli Bolaños-Martínez
2   Laboratorio de Biofarmacéuticos Recombinantes, Universidad Autónoma de San Luis Potosí, SLP, Mexico
,
Andrea Romero Maldonado
2   Laboratorio de Biofarmacéuticos Recombinantes, Universidad Autónoma de San Luis Potosí, SLP, Mexico
3   Sección de Biotecnología, Centro de Investigación en Ciencias de la Salud y Biomedicina (CICSaB), Universidad Autónoma de San Luis Potosí, Mexico
,
Dania O. Govea-Alonso
2   Laboratorio de Biofarmacéuticos Recombinantes, Universidad Autónoma de San Luis Potosí, SLP, Mexico
3   Sección de Biotecnología, Centro de Investigación en Ciencias de la Salud y Biomedicina (CICSaB), Universidad Autónoma de San Luis Potosí, Mexico
,
Christian Carreño-Campos
1   Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, Mexico
,
María Luisa Villarreal
1   Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, Mexico
,
Sergio Rosales-Mendoza
2   Laboratorio de Biofarmacéuticos Recombinantes, Universidad Autónoma de San Luis Potosí, SLP, Mexico
3   Sección de Biotecnología, Centro de Investigación en Ciencias de la Salud y Biomedicina (CICSaB), Universidad Autónoma de San Luis Potosí, Mexico
,
Anabel Ortiz-Caltempa
1   Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, Mexico
› Author AffiliationsThis study was funded by CONACYT/México (grant CF-2019-848290).
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Abstract

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.

Key words

Daucus carota - airlift bioreactor - suspension culture - poliovirus - Umbelliferae - VP2 protein

Supporting Information

    The aspect of cell cultures in solid medium, flask, and airlift bioreactor is presented in Fig. S1 available in the Supporting information.

  • Supporting Information


Publication History

Received: 01 June 2023

Accepted after revision: 20 September 2023

Article published online:
18 October 2023

© 2023. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

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