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Serially subcultivated cells as substrates for poliovirus production for vaccine

A von Seefried, J H Chun

    Developments in Biological Standardization
    |January 1, 1981
    PubMed

    Insights

    This study evaluated human diploid and simian aneuploid cell cultures as alternatives for Inactivated Poliomyelitis Vaccine (IPV) production. Optimizing media composition and utilizing microcarrier systems significantly enhanced viral antigen yields, showing promise for IPV manufacturing.

    Area of Science:

    • Biotechnology and Bioprocessing
    • Vaccine Production
    • Cell Culture Technology

    Background:

    • Primary monkey kidney cells have historically been used for Inactivated Poliomyelitis Vaccine (IPV) production.
    • There is a need for alternative, scalable, and reproducible cell substrates for vaccine manufacturing.
    • Cell culture media composition critically influences cell growth and viral antigen yield.

    Purpose of the Study:

    • To assess human diploid cell lines (WI-38, MRC-5, HEL 299, IMR-90) and a simian aneuploid cell line (LLC-MK2) as alternatives to primary monkey kidney cells for IPV production.
    • To investigate the impact of basal media composition on cell and viral antigen yields.
    • To evaluate the suitability of microcarrier suspension culture systems for IPV manufacturing using these cell substrates.

    Main Methods:

    • Cultured four human diploid cell lines and one simian aneuploid cell line.
    • Optimized basal media (CMRL-1969 and Medium 199 with specific BSS) for cell growth and viral replication.
    • Utilized Multi-Surface-Cell-Propagator (MSCP) and microcarrier suspension culture systems.
    • Measured D-antigen yields for poliovirus Types 1, 2, and 3.

    Main Results:

    • Increased concentrations of specific amino acids in Medium CMRL-1969 significantly improved cell and viral antigen yields.
    • Medium 199 with appropriate BSS enhanced D-antigen yields compared to other media lacking nucleic acid precursors.
    • Microcarrier culture systems achieved 3-4 cell population doublings, with average D-antigen yields of 110 (Type 1), 25 (Type 2), and 35 (Type 3).
    • Human diploid cell cultures exhibited similar physiological requirements, allowing for interchangeability.
    • LLC-MK2 cells demonstrated a reduced serum requirement, a key advantage.

    Conclusions:

    • Human diploid cell cultures (WI-38, MRC-5, HEL 299, IMR-90) and LLC-MK2 cells are viable alternatives for Inactivated Poliomyelitis Vaccine (IPV) production.
    • Optimized media formulations and microcarrier suspension culture systems are crucial for achieving high viral antigen yields.
    • The microcarrier system is suitable for IPV production with these cell substrates, provided optimal suspension is maintained.

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