Effect of different freezing rates during cryopreservation of rat mesenchymal stem cells using combinations of hydroxyethyl starch and dimethylsulfoxide

Background: Mesenchymal stem cells (MSCs) are increasingly used as therapeutic agents as well as research tools in regenerative medicine. Development of technologies which allow storing and banking of MSC with minimal loss of cell viability, differentiation capacity, and function is required for cli...

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Main Authors:	Yahaira Naaldijk, Marek Staude, Viktoriya Fedorova, Alexandra Stolzing
Format:	Default Article
Published:	2012
Subjects:	Mechanical engineering not elsewhere classified Mesenchymal stem cells Cryopreservation Controlled rate freezing Hydroxyethyl starch Mechanical Engineering not elsewhere classified
Online Access:	https://hdl.handle.net/2134/16430
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id	rr-article-9562553
record_format	Figshare
spelling	rr-article-95625532012-01-01T00:00:00Z Effect of different freezing rates during cryopreservation of rat mesenchymal stem cells using combinations of hydroxyethyl starch and dimethylsulfoxide Yahaira Naaldijk (3933095) Marek Staude (5745389) Viktoriya Fedorova (7205159) Alexandra Stolzing (1251348) Mechanical engineering not elsewhere classified Mesenchymal stem cells Cryopreservation Controlled rate freezing Hydroxyethyl starch Mechanical Engineering not elsewhere classified Background: Mesenchymal stem cells (MSCs) are increasingly used as therapeutic agents as well as research tools in regenerative medicine. Development of technologies which allow storing and banking of MSC with minimal loss of cell viability, differentiation capacity, and function is required for clinical and research applications. Cryopreservation is the most effective way to preserve cells long term, but it involves potentially cytotoxic compounds and processing steps. Here, we investigate the effect of decreasing dimethyl sulfoxide (DMSO) concentrations in cryosolution by substituting with hydroxyethyl starch (HES) of different molecular weights using different freezing rates. Post-thaw viability, phenotype and osteogenic differentiation capacity of MSCs were analysed.Results: The study confirms that, for rat MSC, cryopreservation effects need to be assessed some time after, rather than immediately after thawing. MSCs cryopreserved with HES maintain their characteristic cell surface marker expression as well as the osteogenic, adipogenic and chondrogenic differentiation potential. HES alone does not provide sufficient cryoprotection for rat MSCs, but provides good cryoprotection in combination with DMSO, permitting the DMSO content to be reduced to 5%. There are indications that such a combination would seem useful not just for the clinical disadvantages of DMSO but also based on a tendency for reduced osteogenic differentiation capacity of rat MSC cryopreserved with high DMSO concentration. HES molecular weight appears to play only a minor role in its capacity to act as a cryopreservation solution for MSC. The use of a 'straight freeze' protocol is no less effective in maintaining post-thaw viability of MSC compared to controlled rate freezing methods.Conclusion: A 5% DMSO / 5% HES solution cryopreservation solution using a 'straight freeze' approach can be recommended for rat MSC. © 2012 Naaldjik et al.; licensee BioMed Central Ltd. 2012-01-01T00:00:00Z Text Journal contribution 2134/16430 https://figshare.com/articles/journal_contribution/Effect_of_different_freezing_rates_during_cryopreservation_of_rat_mesenchymal_stem_cells_using_combinations_of_hydroxyethyl_starch_and_dimethylsulfoxide/9562553 CC BY 3.0
institution	Loughborough University
collection	Figshare
topic	Mechanical engineering not elsewhere classified Mesenchymal stem cells Cryopreservation Controlled rate freezing Hydroxyethyl starch Mechanical Engineering not elsewhere classified
spellingShingle	Mechanical engineering not elsewhere classified Mesenchymal stem cells Cryopreservation Controlled rate freezing Hydroxyethyl starch Mechanical Engineering not elsewhere classified Yahaira Naaldijk Marek Staude Viktoriya Fedorova Alexandra Stolzing Effect of different freezing rates during cryopreservation of rat mesenchymal stem cells using combinations of hydroxyethyl starch and dimethylsulfoxide
description	Background: Mesenchymal stem cells (MSCs) are increasingly used as therapeutic agents as well as research tools in regenerative medicine. Development of technologies which allow storing and banking of MSC with minimal loss of cell viability, differentiation capacity, and function is required for clinical and research applications. Cryopreservation is the most effective way to preserve cells long term, but it involves potentially cytotoxic compounds and processing steps. Here, we investigate the effect of decreasing dimethyl sulfoxide (DMSO) concentrations in cryosolution by substituting with hydroxyethyl starch (HES) of different molecular weights using different freezing rates. Post-thaw viability, phenotype and osteogenic differentiation capacity of MSCs were analysed.Results: The study confirms that, for rat MSC, cryopreservation effects need to be assessed some time after, rather than immediately after thawing. MSCs cryopreserved with HES maintain their characteristic cell surface marker expression as well as the osteogenic, adipogenic and chondrogenic differentiation potential. HES alone does not provide sufficient cryoprotection for rat MSCs, but provides good cryoprotection in combination with DMSO, permitting the DMSO content to be reduced to 5%. There are indications that such a combination would seem useful not just for the clinical disadvantages of DMSO but also based on a tendency for reduced osteogenic differentiation capacity of rat MSC cryopreserved with high DMSO concentration. HES molecular weight appears to play only a minor role in its capacity to act as a cryopreservation solution for MSC. The use of a 'straight freeze' protocol is no less effective in maintaining post-thaw viability of MSC compared to controlled rate freezing methods.Conclusion: A 5% DMSO / 5% HES solution cryopreservation solution using a 'straight freeze' approach can be recommended for rat MSC. © 2012 Naaldjik et al.; licensee BioMed Central Ltd.
format	Default Article
author	Yahaira Naaldijk Marek Staude Viktoriya Fedorova Alexandra Stolzing
author_facet	Yahaira Naaldijk Marek Staude Viktoriya Fedorova Alexandra Stolzing
author_sort	Yahaira Naaldijk (3933095)
title	Effect of different freezing rates during cryopreservation of rat mesenchymal stem cells using combinations of hydroxyethyl starch and dimethylsulfoxide
title_short	Effect of different freezing rates during cryopreservation of rat mesenchymal stem cells using combinations of hydroxyethyl starch and dimethylsulfoxide
title_full	Effect of different freezing rates during cryopreservation of rat mesenchymal stem cells using combinations of hydroxyethyl starch and dimethylsulfoxide
title_fullStr	Effect of different freezing rates during cryopreservation of rat mesenchymal stem cells using combinations of hydroxyethyl starch and dimethylsulfoxide
title_full_unstemmed	Effect of different freezing rates during cryopreservation of rat mesenchymal stem cells using combinations of hydroxyethyl starch and dimethylsulfoxide
title_sort	effect of different freezing rates during cryopreservation of rat mesenchymal stem cells using combinations of hydroxyethyl starch and dimethylsulfoxide
publishDate	2012
url	https://hdl.handle.net/2134/16430
_version_	1797558640968728576

Effect of different freezing rates during cryopreservation of rat mesenchymal stem cells using combinations of hydroxyethyl starch and dimethylsulfoxide

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