Development of a serum-free medium to aid large-scale production of mycoplasma-based therapies
Autor/a
Burgos, Raul
Garcia-Ramallo, Eva
Shaw, Daniel
Lluch-Senar, Maria
Serrano, Luis
Data de publicació
2023ISSN
2165-0497
Resum
To assist in the advancement of the large-scale production of safe Mycoplasma vaccines and other Mycoplasma-based therapies, we developed a culture medium free of animal serum and other animal components for Mycoplasma pneumoniae growth. By establishing a workflow method to systematically test different compounds and concentrations, we provide optimized formulations capable of supporting serial passaging and robust growth reaching 60 to 70% of the biomass obtained in rich medium. Global transcriptomic and proteomic analysis showed minor physiological changes upon cell culture in the animal component-free medium, supporting its suitability for the production of M. pneumoniae-based therapies. The major contributors to growth performance were found to be glucose as a carbon source, glycerol, cholesterol, and phospholipids as a source of fatty acids. Bovine serum albumin or cyclodextrin (in the animal component-free medium) were required as lipid carriers to prevent lipid toxicity. Connaught Medical Research Laboratories medium (CMRL) used to simplify medium preparation as a source of amino acids, nucleotide precursors, vitamins, and other cofactors could be substituted by cysteine. In fact, the presence of protein hydrolysates such as yeastolate or peptones was found to be essential and preferred over free amino acids, except for the cysteine. Supplementation of nucleotide precursors and vitamins is not strictly necessary in the presence of yeastolate, suggesting that this animal origin-free hydrolysate serves as an efficient source for these compounds. Finally, we adapted the serum-free medium formulation to support growth of Mycoplasma hyopneumoniae, a swine pathogen for which inactivated whole-cell vaccines are available.
Tipus de document
Article
Versió del document
Versió publicada
Llengua
English
Matèries (CDU)
61 - Medicina
Paraules clau
Micoplasma
Medis
Metabolisme
Medi sense sèrum
Vacunes
Micoplasma
Medios
Metabolismo
Medio libre de suero
Vacunas
Mycoplasma
Media
Metabolisme
Serum-free medium
Vaccines
Pàgines
14
Publicat per
American Society for Microbiology
Col·lecció
11; 3
Publicat a
Microbiology Spectrum
Citació
Burgos, Raul; Garcia-Ramallo, Eva; Shaw, Daniel [et al.]. Development of a serum-free medium to aid large-scale production of mycoplasma-based therapies. Microbiology Spectrum, 2023, 11(3), p. 1-14. Disponible en: <https://journals.asm.org/doi/10.1128/spectrum.04859-22>. Fecha de acceso: 17 oct. 2023. DOI: 10.1128/spectrum.04859-22
Aquest element apareix en la col·lecció o col·leccions següent(s)
- Ciències de la Salut [745]
Drets
© 2023 Burgos et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license
Excepte que s'indiqui una altra cosa, la llicència de l'ítem es descriu com https://creativecommons.org/licenses/by/4.0/