Significance of nutrient media choice for the long-term cultures of leukemic T-lymphoblasts

Correct choice of nutrient media for culturing different types of cells in various applications is one of the most important aspects of modern biotechnology, since chemical composition of the culture media largely contains the necessary metabolites to support certain cells’ growth lines outside the...

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Autores principales:	L. S. Litvinova, K. A. Yurova, V. V. Shchupletsova, N. D. Gazatova, O. G. Khaziakhmatova, V. V. Malashchenko, E. O. Shunkin, N. M. Todosenko, E. S. Melashchenko, M. Yu. Khlusova, I. А. Khlusov
Formato:	article
Lenguaje:	RU
Publicado:	SPb RAACI 2021
Materias:	rpmi 1640 amem jurkat t cells viability secretion cell-iq monitoring Immunologic diseases. Allergy RC581-607
Acceso en línea:	https://doaj.org/article/4812ab5472be485f8ae7fffc982c7905
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spelling	oai:doaj.org-article:4812ab5472be485f8ae7fffc982c79052021-11-18T08:03:51ZSignificance of nutrient media choice for the long-term cultures of leukemic T-lymphoblasts1563-06252313-741X10.15789/1563-0625-SON-2142https://doaj.org/article/4812ab5472be485f8ae7fffc982c79052021-06-01T00:00:00Zhttps://www.mimmun.ru/mimmun/article/view/2142https://doaj.org/toc/1563-0625https://doaj.org/toc/2313-741XCorrect choice of nutrient media for culturing different types of cells in various applications is one of the most important aspects of modern biotechnology, since chemical composition of the culture media largely contains the necessary metabolites to support certain cells’ growth lines outside the body. Jurkat line of human leukemic T-lymphoblast-like cells (hereinafter Jurkat T-cells) is actively used for in vitro modeling of intracellular signaling and activation of normal blood T-lymphocytes mediated by the T-cell receptor/CD3/ CD4 complex in toxicological studies of immune and secretory responses, to test medicinal substances and ions. Also, Jurkat T-cells are widely used for ex vivo testing in immunology, oncology, toxicology, orthopedics, and traumatology. The existing standards and numerous studies are mainly based on short-term in vitro cultivation of Jurkat T-cells in RPMI 1640 nutrient medium. Meanwhile, the issues of long-term maintenance of the growth of Jurkat T-cells culture are poorly presented in the research literature. This study aimed for studying the activity of Jurkat T-cells over 7 to 14 days of in vitro culture and comparing the relative value of RPMI 1640 and αMEM media for the behavior of immunocompetent tumor cells. Using flow cytometry, multiplex analysis, and phase contrast Cell-IQ microscopy, the proportions of living cells and those dying by apoptosis and necrosis, secretion of cytokines and chemokines, and the dynamics of cell biomass propagation were studied. It was found that the αMEM medium in the complete nutrient medium, as compared with RPMI 1640, is more appropriate to in vitro promotion of cell viability (increased proportion of viable cells by 13.5% at the day 14), their secretory ability for 23 из 27 tested biomolecules, shortened adaptation time (на 32%) in culture before growth initiation, 5-fold increase of the Jurkat Т-cell cellularity by the day 7. Potential significance of the chemical components of nutrient media and secreted biomolecules for these results is discussed. As based on the results obtained, we concluded on superior properties of αMEM medium for long-term in vitro cultures of Jurkat T-cells. Consequently, the in vitro testing of medical devices intended for long-term contact with the body, including those for cancer patients, using Jurkat T-cell leukemia line in RPMI 1640 medium, may lead to wrong predictions on their biocompatibility and potential antitumor activity.L. S. LitvinovaK. A. YurovaV. V. ShchupletsovaN. D. GazatovaO. G. KhaziakhmatovaV. V. MalashchenkoE. O. ShunkinN. M. TodosenkoE. S. MelashchenkoM. Yu. KhlusovaI. А. KhlusovSPb RAACIarticlerpmi 1640amemjurkat t cellsviabilitysecretioncell-iq monitoringImmunologic diseases. AllergyRC581-607RUMedicinskaâ Immunologiâ, Vol 23, Iss 3, Pp 593-604 (2021)
institution	DOAJ
collection	DOAJ
language	RU
topic	rpmi 1640 amem jurkat t cells viability secretion cell-iq monitoring Immunologic diseases. Allergy RC581-607
spellingShingle	rpmi 1640 amem jurkat t cells viability secretion cell-iq monitoring Immunologic diseases. Allergy RC581-607 L. S. Litvinova K. A. Yurova V. V. Shchupletsova N. D. Gazatova O. G. Khaziakhmatova V. V. Malashchenko E. O. Shunkin N. M. Todosenko E. S. Melashchenko M. Yu. Khlusova I. А. Khlusov Significance of nutrient media choice for the long-term cultures of leukemic T-lymphoblasts
description	Correct choice of nutrient media for culturing different types of cells in various applications is one of the most important aspects of modern biotechnology, since chemical composition of the culture media largely contains the necessary metabolites to support certain cells’ growth lines outside the body. Jurkat line of human leukemic T-lymphoblast-like cells (hereinafter Jurkat T-cells) is actively used for in vitro modeling of intracellular signaling and activation of normal blood T-lymphocytes mediated by the T-cell receptor/CD3/ CD4 complex in toxicological studies of immune and secretory responses, to test medicinal substances and ions. Also, Jurkat T-cells are widely used for ex vivo testing in immunology, oncology, toxicology, orthopedics, and traumatology. The existing standards and numerous studies are mainly based on short-term in vitro cultivation of Jurkat T-cells in RPMI 1640 nutrient medium. Meanwhile, the issues of long-term maintenance of the growth of Jurkat T-cells culture are poorly presented in the research literature. This study aimed for studying the activity of Jurkat T-cells over 7 to 14 days of in vitro culture and comparing the relative value of RPMI 1640 and αMEM media for the behavior of immunocompetent tumor cells. Using flow cytometry, multiplex analysis, and phase contrast Cell-IQ microscopy, the proportions of living cells and those dying by apoptosis and necrosis, secretion of cytokines and chemokines, and the dynamics of cell biomass propagation were studied. It was found that the αMEM medium in the complete nutrient medium, as compared with RPMI 1640, is more appropriate to in vitro promotion of cell viability (increased proportion of viable cells by 13.5% at the day 14), their secretory ability for 23 из 27 tested biomolecules, shortened adaptation time (на 32%) in culture before growth initiation, 5-fold increase of the Jurkat Т-cell cellularity by the day 7. Potential significance of the chemical components of nutrient media and secreted biomolecules for these results is discussed. As based on the results obtained, we concluded on superior properties of αMEM medium for long-term in vitro cultures of Jurkat T-cells. Consequently, the in vitro testing of medical devices intended for long-term contact with the body, including those for cancer patients, using Jurkat T-cell leukemia line in RPMI 1640 medium, may lead to wrong predictions on their biocompatibility and potential antitumor activity.
format	article
author	L. S. Litvinova K. A. Yurova V. V. Shchupletsova N. D. Gazatova O. G. Khaziakhmatova V. V. Malashchenko E. O. Shunkin N. M. Todosenko E. S. Melashchenko M. Yu. Khlusova I. А. Khlusov
author_facet	L. S. Litvinova K. A. Yurova V. V. Shchupletsova N. D. Gazatova O. G. Khaziakhmatova V. V. Malashchenko E. O. Shunkin N. M. Todosenko E. S. Melashchenko M. Yu. Khlusova I. А. Khlusov
author_sort	L. S. Litvinova
title	Significance of nutrient media choice for the long-term cultures of leukemic T-lymphoblasts
title_short	Significance of nutrient media choice for the long-term cultures of leukemic T-lymphoblasts
title_full	Significance of nutrient media choice for the long-term cultures of leukemic T-lymphoblasts
title_fullStr	Significance of nutrient media choice for the long-term cultures of leukemic T-lymphoblasts
title_full_unstemmed	Significance of nutrient media choice for the long-term cultures of leukemic T-lymphoblasts
title_sort	significance of nutrient media choice for the long-term cultures of leukemic t-lymphoblasts
publisher	SPb RAACI
publishDate	2021
url	https://doaj.org/article/4812ab5472be485f8ae7fffc982c7905
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Significance of nutrient media choice for the long-term cultures of leukemic T-lymphoblasts

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