Effect of Different Freezing Rates During Cryopreservation of Rat Mesenchymal Stem Cells Using Combinations of Hydroxyethyl Starch and Dimethylsulfoxide

Overview

Journal BMC Biotechnol

Publisher Biomed Central

Specialty Biotechnology

Date 2012 Aug 15

PMID 22889198

Citations 24

Authors

Yahaira Naaldijk

Marek Staude

Viktoriya Fedorova

Alexandra Stolzing

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

Exploring the Potential Effects of Cryopreservation on the Biological Characteristics and Cardiomyogenic Differentiation of Rat Adipose-Derived Mesenchymal Stem Cells.

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Optimizing cryopreservation conditions for use of fucosylated human mesenchymal stromal cells in anti-inflammatory/immunomodulatory therapeutics.

Gil-Chinchilla J, Bueno C, Martinez C, Ferrandez-Murtula A, Garcia-Hernandez A, Blanquer M Front Immunol. 2024; 15:1385691.

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Addition of synthetic polymer in the freezing solution of mesenchymal stem cells from equine adipose tissue as a future perspective for reducing of DMSO concentration.

Nascimento C, Saraiva M, Mattos Pereira V, de Brito D, de Aguiar F, Alves B Braz J Vet Med. 2024; 45:e002523.

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Cryopreservation of pig spermatozoa using carboxylated poly-L-lysine as cryoprotectant.

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hMSCs in contact with DMSO for cryopreservation: Experiments and modeling of osmotic injury and cytotoxic effect.

Traversari G, Delogu F, Aparicio S, Cincotti A Biotechnol Bioeng. 2022; 119(10):2890-2907.

PMID: 35799310 PMC: 9546233. DOI: 10.1002/bit.28174.

References

Woodbury D, Schwarz E, Prockop D, Black I . Adult rat and human bone marrow stromal cells differentiate into neurons. J Neurosci Res. 2000; 61(4):364-70. DOI: 10.1002/1097-4547(20000815)61:4<364::AID-JNR2>3.0.CO;2-C. View

Moon J, Kwak S, Park G, Jung H, Yoon B, Park J . Isolation and characterization of multipotent human keloid-derived mesenchymal-like stem cells. Stem Cells Dev. 2008; 17(4):713-24. DOI: 10.1089/scd.2007.0210. View

Thirumala S, Gimble J, Devireddy R . Evaluation of methylcellulose and dimethyl sulfoxide as the cryoprotectants in a serum-free freezing media for cryopreservation of adipose-derived adult stem cells. Stem Cells Dev. 2009; 19(4):513-22. PMC: 3139530. DOI: 10.1089/scd.2009.0173. View

Sputtek A, Jetter S, Hummel K, Kuhnl P . Cryopreservation of peripheral blood progenitor cells: characteristics of suitable techniques. Beitr Infusionsther Transfusionsmed. 1997; 34:79-83. View

Westphal M, Lamszus K . Other experimental therapies for glioma. Recent Results Cancer Res. 2009; 171:155-64. DOI: 10.1007/978-3-540-31206-2_9. View

Ashwood-Smith M, Warby C, Connor K, Becker G . Low-temperature preservation of mammalian cells in tissue culture with polyvinylpyrrolidone (PVP), dextrans, and hydroxyethyl starch (HES). Cryobiology. 1972; 9(5):441-9. DOI: 10.1016/0011-2240(72)90161-7. View

Reyes M, Verfaillie C . Characterization of multipotent adult progenitor cells, a subpopulation of mesenchymal stem cells. Ann N Y Acad Sci. 2001; 938:231-3; discussion 233-5. DOI: 10.1111/j.1749-6632.2001.tb03593.x. View

Liu G, Zhou H, Li Y, Li G, Cui L, Liu W . Evaluation of the viability and osteogenic differentiation of cryopreserved human adipose-derived stem cells. Cryobiology. 2008; 57(1):18-24. DOI: 10.1016/j.cryobiol.2008.04.002. View

Wakitani S, Saito T, Caplan A . Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-azacytidine. Muscle Nerve. 1995; 18(12):1417-26. DOI: 10.1002/mus.880181212. View

10.

Carvalho K, Cury C, Oliveira L, Cattaned R, Malvezzi M, Francisco J . Evaluation of bone marrow mesenchymal stem cell standard cryopreservation procedure efficiency. Transplant Proc. 2008; 40(3):839-41. DOI: 10.1016/j.transproceed.2008.03.004. View

11.

Liu Y, Xu X, Ma X, Liu J, Cui Z . Effect of various freezing solutions on cryopreservation of mesenchymal stem cells from different animal species. Cryo Letters. 2011; 32(5):425-35. View

12.

Kandra D, Devireddy R . Numerical Simulation of Local Temperature Distortions During Ice Nucleation of Cells in Suspension. Int J Heat Mass Transf. 2011; 51(23-24):5655-5661. PMC: 3147025. DOI: 10.1016/j.ijheatmasstransfer.2008.04.026. View

13.

Martinello T, Bronzini I, Maccatrozzo L, Mollo A, Sampaolesi M, Mascarello F . Canine adipose-derived-mesenchymal stem cells do not lose stem features after a long-term cryopreservation. Res Vet Sci. 2010; 91(1):18-24. DOI: 10.1016/j.rvsc.2010.07.024. View

14.

Reubinoff B, Pera M, Vajta G, Trounson A . Effective cryopreservation of human embryonic stem cells by the open pulled straw vitrification method. Hum Reprod. 2001; 16(10):2187-94. DOI: 10.1093/humrep/16.10.2187. View

15.

Perry B, Zhou D, Wu X, Yang F, Byers M, Chu T . Collection, cryopreservation, and characterization of human dental pulp-derived mesenchymal stem cells for banking and clinical use. Tissue Eng Part C Methods. 2008; 14(2):149-56. PMC: 2963629. DOI: 10.1089/ten.tec.2008.0031. View

16.

Teasdale B, Sieber V, Riches D, Nanchahal J . Cryopreservation of cultured dermal fibroblast impregnated collagen gels. Burns. 1993; 19(5):406-10. DOI: 10.1016/0305-4179(93)90062-d. View

17.

Lee R, Kim B, Choi I, Kim H, Choi H, Suh K . Characterization and expression analysis of mesenchymal stem cells from human bone marrow and adipose tissue. Cell Physiol Biochem. 2004; 14(4-6):311-24. DOI: 10.1159/000080341. View

18.

Baust J, Van Buskirk , Baust J . Cell viability improves following inhibition of cryopreservation-induced apoptosis. In Vitro Cell Dev Biol Anim. 2000; 36(4):262-70. DOI: 10.1290/1071-2690(2000)036<0262:cvifio>2.0.co;2. View

19.

Stiff P, Koester A, Weidner M, Dvorak K, Fisher R . Autologous bone marrow transplantation using unfractionated cells cryopreserved in dimethylsulfoxide and hydroxyethyl starch without controlled-rate freezing. Blood. 1987; 70(4):974-8. View

20.

Bhakta G, Lee K, Magalhaes R, Wen F, Gouk S, Hutmacher D . Cryopreservation of alginate-fibrin beads involving bone marrow derived mesenchymal stromal cells by vitrification. Biomaterials. 2008; 30(3):336-43. DOI: 10.1016/j.biomaterials.2008.09.030. View