A CAR/CD19 Jurkat cell model for bioprocess screening and expansion: potential use for CAR-T cells production.
6 affiliations
- 1Pharmaceutical Biotechnology Laboratory, Faculty of Pharmacy, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, Mexico.
- 2Secretaría de Ciencia, Humanidades, Tecnología e Innovación, SECIHTI, Mexico City, Mexico.
- 3Comisión Coordinadora de Institutos Nacionales de Salud y Hospitales de Alta Especialidad (CCINSHAE), Ministry of Health (Secretaría de Salud), Mexico City, Mexico.
- 4Immunodeficiencies Research Laboratory and Clinical Immunology Department, Instituto Nacional de Pediatría, Mexico City, Mexico.
- 5Department of Research and Development, Triovance SAPI de CV, Mexico City, Mexico.
- 6Hematopoietic Stem Cell Transplantation and Cell Therapy Program, Instituto Nacional de Pediatría, Mexico City, Mexico.
Abstract
Introduction
Cell expansion is one of the most critical steps in the development of CAR-T cell manufacturing processes because it directly influences cell yield and process performance. The systematic optimization of expansion parameters using primary human T cells is often limited by inter-donor variability, cost, and experimental complexity. So, it is necessary to study more homogeneous populations that allow to stablish standardized conditions in a controlled environment that were able to be used as standard. So, in this study, we developed a Jurkat CAR-CD19 cell-based platform as a reproducible model for optimizing the cell expansion bioprocess under controlled culture conditions.Methods
CAR-CD19-expressing Jurkat cells were pre-generated, molecularly characterized and subsequently used to evaluate key bioprocess variables, including glucose concentration, inoculum density, culture medium composition (RPMI + 10% FBS versus Nutri-T without serum), shaking speed, and different culture operating modes (batch, fed-batch, and semi-perfusion). Cell growth kinetics, metabolite consumption, metabolite production, and CAR expression were evaluated throughout the optimization process.Results
An intermediate glucose concentration supported cell expansion without additional benefit from higher glucose supplementation. Serum-free Nutri-T medium promoted higher viable cell densities than RPMI (used as control media) under the evaluated conditions. Among the operating modes evaluated, semi-perfusion achieved the highest viable cell density, maintaining high cell viability and a homogeneous CAR-positive cell population at the end of culture in Nutri-T media.Conclusion
These findings demonstrate that the Jurkat CAR-CD19 model provides a robust and reproducible experimental platform for optimizing cell expansion bioprocesses. While further validation in primary human CAR-T cells is required, this model allows for the systematic evaluation of culture variables, minimizing experimental variability and facilitating early-stage process development for adoptive cell therapies.
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BibTeX
@article{fuentesbustos2026carcd19,
title = {{A CAR/CD19 Jurkat cell model for bioprocess screening and expansion: potential use for CAR-T cells production.}},
author = {Hugo E Fuentes-Bustos and Alejandrina Hernández-López and Marta Zapata-Tarrés and Juan Carlos Bustamante-Ogando and Blanca Angélica Segura-Pacheco and Alberto Olaya-Vargas and Angélica Meneses-Acosta},
journal = {Frontiers in bioengineering and biotechnology},
year = {2026},
month = oct,
doi = {10.3389/fbioe.2026.1917849},
url = {https://europepmc.org/article/MED/42909135},
note = {CC BY},
}
RIS
TY - JOUR
TI - A CAR/CD19 Jurkat cell model for bioprocess screening and expansion: potential use for CAR-T cells production.
AU - Hugo E Fuentes-Bustos
AU - Alejandrina Hernández-López
AU - Marta Zapata-Tarrés
AU - Juan Carlos Bustamante-Ogando
AU - Blanca Angélica Segura-Pacheco
AU - Alberto Olaya-Vargas
AU - Angélica Meneses-Acosta
PY - 2026
DA - 2026/10/07
JO - Frontiers in bioengineering and biotechnology
DO - 10.3389/fbioe.2026.1917849
UR - https://europepmc.org/article/MED/42909135
AB - Introduction Cell expansion is one of the most critical steps in the development of CAR-T cell manufacturing processes because it directly influences cell yield and process performance. The systematic optimization of expansion parameters using primary human T cells is often limited by inter-donor variability, cost, and experimental complexity. So, it is necessary to study more homogeneous populations that allow to stablish standardized conditions in a controlled environment that were able to be used as standard. So, in this study, we developed a Jurkat CAR-CD19 cell-based platform as a reproducible model for optimizing the cell expansion bioprocess under controlled culture conditions.Methods CAR-CD19-expressing Jurkat cells were pre-generated, molecularly characterized and subsequently used to evaluate key bioprocess variables, including glucose concentration, inoculum density, culture medium composition (RPMI + 10% FBS versus Nutri-T without serum), shaking speed, and different culture operating modes (batch, fed-batch, and semi-perfusion). Cell growth kinetics, metabolite consumption, metabolite production, and CAR expression were evaluated throughout the optimization process.Results An intermediate glucose concentration supported cell expansion without additional benefit from higher glucose supplementation. Serum-free Nutri-T medium promoted higher viable cell densities than RPMI (used as control media) under the evaluated conditions. Among the operating modes evaluated, semi-perfusion achieved the highest viable cell density, maintaining high cell viability and a homogeneous CAR-positive cell population at the end of culture in Nutri-T media.Conclusion These findings demonstrate that the Jurkat CAR-CD19 model provides a robust and reproducible experimental platform for optimizing cell expansion bioprocesses. While further validation in primary human CAR-T cells is required, this model allows for the systematic evaluation of culture variables, minimizing experimental variability and facilitating early-stage process development for adoptive cell therapies.
SN - 2296-4185
ER -