Freezing Medium Containing 5% DMSO Enhances the Cell Viability and Recovery Rate After Cryopreservation of Regulatory T Cell Products ex vivo and in vivo

Cell therapies have significant therapeutic potential in diverse fields including regenerative medicine, transplantation tolerance, and autoimmunity. Within these fields, regulatory T cells (Treg) have been deployed to ameliorate aberrant immune responses with great success. However, translation of...

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Autores principales:	Daniel Kaiser, Natalie Maureen Otto, Oliver McCallion, Henrike Hoffmann, Ghazaleh Zarrinrad, Maik Stein, Carola Beier, Isabell Matz, Marleen Herschel, Joanna Hester, Guido Moll, Fadi Issa, Petra Reinke, Andy Roemhild
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	cell therapy regulatory T cells (Tregs) cryopreservation freeze-thawing freezing medium cell recovery rate Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/50e3a4d1a00a4962920bd8d701178c0c
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spelling	oai:doaj.org-article:50e3a4d1a00a4962920bd8d701178c0c2021-12-03T05:04:31ZFreezing Medium Containing 5% DMSO Enhances the Cell Viability and Recovery Rate After Cryopreservation of Regulatory T Cell Products ex vivo and in vivo2296-634X10.3389/fcell.2021.750286https://doaj.org/article/50e3a4d1a00a4962920bd8d701178c0c2021-12-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fcell.2021.750286/fullhttps://doaj.org/toc/2296-634XCell therapies have significant therapeutic potential in diverse fields including regenerative medicine, transplantation tolerance, and autoimmunity. Within these fields, regulatory T cells (Treg) have been deployed to ameliorate aberrant immune responses with great success. However, translation of the cryopreservation strategies employed for other cell therapy products, such as effector T cell therapies, to Treg therapies has been challenging. The lack of an optimized cryopreservation strategy for Treg products presents a substantial obstacle to their broader application, particularly as administration of fresh cells limits the window available for sterility and functional assessment. In this study, we aimed to develop an optimized cryopreservation strategy for our CD4+CD25+Foxp3+ Treg clinical product. We investigate the effect of synthetic or organic cryoprotectants including different concentrations of DMSO on Treg recovery, viability, phenotype, cytokine production, suppressive capacity, and in vivo survival following GMP-compliant manufacture. We additionally assess the effect of adding the extracellular cryoprotectant polyethylene glycol (PEG), or priming cellular expression of heat shock proteins as strategies to improve viability. We find that cryopreservation in serum-free freezing medium supplemented with 10% human serum albumin and 5% DMSO facilitates improved Treg recovery and functionality and supports a reduced DMSO concentration in Treg cryopreservation protocols. This strategy may be easily incorporated into clinical manufacture protocols for future studies.Daniel KaiserDaniel KaiserNatalie Maureen OttoNatalie Maureen OttoOliver McCallionHenrike HoffmannHenrike HoffmannGhazaleh ZarrinradMaik SteinMaik SteinCarola BeierCarola BeierIsabell MatzMarleen HerschelJoanna HesterGuido MollFadi IssaPetra ReinkePetra ReinkePetra ReinkeAndy RoemhildAndy RoemhildFrontiers Media S.A.articlecell therapyregulatory T cells (Tregs)cryopreservationfreeze-thawingfreezing mediumcell recovery rateBiology (General)QH301-705.5ENFrontiers in Cell and Developmental Biology, Vol 9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	cell therapy regulatory T cells (Tregs) cryopreservation freeze-thawing freezing medium cell recovery rate Biology (General) QH301-705.5
spellingShingle	cell therapy regulatory T cells (Tregs) cryopreservation freeze-thawing freezing medium cell recovery rate Biology (General) QH301-705.5 Daniel Kaiser Daniel Kaiser Natalie Maureen Otto Natalie Maureen Otto Oliver McCallion Henrike Hoffmann Henrike Hoffmann Ghazaleh Zarrinrad Maik Stein Maik Stein Carola Beier Carola Beier Isabell Matz Marleen Herschel Joanna Hester Guido Moll Fadi Issa Petra Reinke Petra Reinke Petra Reinke Andy Roemhild Andy Roemhild Freezing Medium Containing 5% DMSO Enhances the Cell Viability and Recovery Rate After Cryopreservation of Regulatory T Cell Products ex vivo and in vivo
description	Cell therapies have significant therapeutic potential in diverse fields including regenerative medicine, transplantation tolerance, and autoimmunity. Within these fields, regulatory T cells (Treg) have been deployed to ameliorate aberrant immune responses with great success. However, translation of the cryopreservation strategies employed for other cell therapy products, such as effector T cell therapies, to Treg therapies has been challenging. The lack of an optimized cryopreservation strategy for Treg products presents a substantial obstacle to their broader application, particularly as administration of fresh cells limits the window available for sterility and functional assessment. In this study, we aimed to develop an optimized cryopreservation strategy for our CD4+CD25+Foxp3+ Treg clinical product. We investigate the effect of synthetic or organic cryoprotectants including different concentrations of DMSO on Treg recovery, viability, phenotype, cytokine production, suppressive capacity, and in vivo survival following GMP-compliant manufacture. We additionally assess the effect of adding the extracellular cryoprotectant polyethylene glycol (PEG), or priming cellular expression of heat shock proteins as strategies to improve viability. We find that cryopreservation in serum-free freezing medium supplemented with 10% human serum albumin and 5% DMSO facilitates improved Treg recovery and functionality and supports a reduced DMSO concentration in Treg cryopreservation protocols. This strategy may be easily incorporated into clinical manufacture protocols for future studies.
format	article
author	Daniel Kaiser Daniel Kaiser Natalie Maureen Otto Natalie Maureen Otto Oliver McCallion Henrike Hoffmann Henrike Hoffmann Ghazaleh Zarrinrad Maik Stein Maik Stein Carola Beier Carola Beier Isabell Matz Marleen Herschel Joanna Hester Guido Moll Fadi Issa Petra Reinke Petra Reinke Petra Reinke Andy Roemhild Andy Roemhild
author_facet	Daniel Kaiser Daniel Kaiser Natalie Maureen Otto Natalie Maureen Otto Oliver McCallion Henrike Hoffmann Henrike Hoffmann Ghazaleh Zarrinrad Maik Stein Maik Stein Carola Beier Carola Beier Isabell Matz Marleen Herschel Joanna Hester Guido Moll Fadi Issa Petra Reinke Petra Reinke Petra Reinke Andy Roemhild Andy Roemhild
author_sort	Daniel Kaiser
title	Freezing Medium Containing 5% DMSO Enhances the Cell Viability and Recovery Rate After Cryopreservation of Regulatory T Cell Products ex vivo and in vivo
title_short	Freezing Medium Containing 5% DMSO Enhances the Cell Viability and Recovery Rate After Cryopreservation of Regulatory T Cell Products ex vivo and in vivo
title_full	Freezing Medium Containing 5% DMSO Enhances the Cell Viability and Recovery Rate After Cryopreservation of Regulatory T Cell Products ex vivo and in vivo
title_fullStr	Freezing Medium Containing 5% DMSO Enhances the Cell Viability and Recovery Rate After Cryopreservation of Regulatory T Cell Products ex vivo and in vivo
title_full_unstemmed	Freezing Medium Containing 5% DMSO Enhances the Cell Viability and Recovery Rate After Cryopreservation of Regulatory T Cell Products ex vivo and in vivo
title_sort	freezing medium containing 5% dmso enhances the cell viability and recovery rate after cryopreservation of regulatory t cell products ex vivo and in vivo
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/50e3a4d1a00a4962920bd8d701178c0c
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Freezing Medium Containing 5% DMSO Enhances the Cell Viability and Recovery Rate After Cryopreservation of Regulatory T Cell Products ex vivo and in vivo

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