Abstract
The capability of multipotent mesenchymal stem cells to maintain cell viability, phenotype and differentiation ability upon thawing is critical if they are to be banked and used for future therapeutic purposes. In the present study, we examined the effect of 9-10 months of cryostorage on the morphology, immunophenotype, colony-forming unit (CFU) and differentiation capacity of fresh and cryopreserved human adipose-derived stromal/stem cells (ASCs) from the same donors. Cryopreservation did not reduce the CFU frequency and the expression levels of CD29, CD73, CD90 and CD105 remained unchanged with the exception of CD34 and CD45; however, the differentiation capacity of cryopreserved ASCs relative to fresh cells was significantly reduced. While our findings suggest that future studies are warranted to improve cryopreservation methods and agents, cryopreserved ASCs retain sufficient features to ensure their practical utility for both research and clinical applications.
References
1.
Bakken, A.M. (2006) Cryopreserving human peripheral blood progenitor cells. Curr Stem Cell Res Ther 1: 47-54.
2.
Bourin, P., B.A. Bunnell, L. Casteilla, M. Dominici, A.J. Katz, K.L. March, H. Redl, J.P. Rubin, K. Yoshimura, J.M. Gimble (2013) Stromal cells from the adipose tissue-derived stromal vascular fraction and culture expanded adipose tissue-derived stromal/stem cells: a joint statement of the International Federation for Adipose Therapeutics and Science (IFATS) and the International Society for Cellular Therapy (ISCT). Cytotherapy 15: 641-648.
3.
Caplan, A.I. (1991) Mesenchymal stem cells. J Orthop Res 9: 641-650.
4.
Carvalho, P.P., X. Wu, G. Yu, M. Dietrich, I.R. Dias, M.E. Gomes, R.L. Reis, J.M. Gimble (2011) Use of animal protein-free products for passaging adherent human adipose-derived stromal/stem cells. Cytotherapy 13: 594-597.
5.
Castro-Malaspina, H., R.E. Gay, S.C. Jhanwar, J.A. Hamilton, D.R. Chiarieri, P.A. Meyers, S. Gay, M.A. Moore (1982) Characteristics of bone marrow fibroblast colony-forming cells (CFU-F) and their progeny in patients with myeloproliferative disorders. Blood 59: 1046-1054.
6.
De Rosa, A., F. De Francesco, V. Tirino, G.A. Ferraro, V. Desiderio, F. Paino, G. Pirozzi, F. D'Andrea, G. Papaccio (2009) A new method for cryopreserving adipose-derived stem cells: an attractive and suitable large-scale and long-term cell banking technology. Tissue Eng Part C Methods 15: 659-667.
7.
Devireddy, R.V., S. Thirumala, J.M. Gimble (2005) Cellular response of adipose derived passage-4 adult stem cells to freezing stress. J Biomech Eng 127: 1081-1086.
8.
Dominici, M., K. Le Blanc, I. Mueller, I. Slaper-Cortenbach, F.C. Marini, D.S. Krause, R.J. Deans, A. Keating, D.J. Prockop, E.M. Horwitz (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8: 315-317.
9.
Fleming, K.K., A. Hubel (2006) Cryopreservation of hematopoietic and non-hematopoietic stem cells. Transfus Apher Sci 34: 309-315.
10.
Gimble, J.M., F. Guilak (2003) Adipose-derived adult stem cells: isolation, characterization, and differentiation potential. Cytotherapy 5: 362-369.
11.
Gonzalez-Fernandez, M.L., S. Perez-Castrillo, P. Ordas-Fernandez, M.E. Lopez-Gonzalez, B. Colaco, V. Villar-Suarez (2015) Study on viability and chondrogenic differentiation of cryopreserved adipose tissue-derived mesenchymal stromal cells for future use in regenerative medicine. Cryobiology 71: 256-263.
12.
Haynesworth, S.E., M.A. Baber, A.I. Caplan (1992) Cell surface antigens on human marrow-derived mesenchymal cells are detected by monoclonal antibodies. Bone 13: 69-80.
13.
Horwitz, E.M., K. Le Blanc, M. Dominici, I. Mueller, I. Slaper-Cortenbach, F.C. Marini, R.J. Deans, D.S. Krause, A. Keating (2005) Clarification of the nomenclature for MSC: The International Society for Cellular Therapy position statement. Cytotherapy 7: 393-395.
14.
James, A.W., B. Levi, E.R. Nelson, M. Peng, G.W. Commons, M. Lee, B. Wu, M.T. Longaker (2011) Deleterious effects of freezing on osteogenic differentiation of human adipose-derived stromal cells in vitro and in vivo. Stem Cells Dev 20: 427-439.
15.
Li, Y., T. Ma (2012) Bioprocessing of cryopreservation for large-scale banking of human pluripotent stem cells. Biores Open Access 1: 205-214.
16.
Liu, G., H. Zhou, Y. Li, G. Li, L. Cui, W. Liu, Y. Cao (2008) Evaluation of the viability and osteogenic differentiation of cryopreserved human adipose-derived stem cells. Cryobiology 57: 18-24.
17.
McIntosh, K., S. Zvonic, S. Garrett, J.B. Mitchell, Z.E. Floyd, L. Hammill, A. Kloster, Y. Di Halvorsen, J.P. Ting, R.W. Storms, B. Goh, G. Kilroy, X. Wu, J.M. Gimble (2006) The immunogenicity of human adipose-derived cells: temporal changes in vitro. Stem Cells 24: 1246-1253.
18.
Mitchell, J.B., K. McIntosh, S. Zvonic, S. Garrett, Z.E. Floyd, A. Kloster, Y. Di Halvorsen, R.W. Storms, B. Goh, G. Kilroy, X. Wu, J.M. Gimble (2006) Immunophenotype of human adipose-derived cells: temporal changes in stromal-associated and stem cell-associated markers. Stem Cells 24: 376-385.
19.
Nakamura, K., S. Nakano, T. Miyoshi, K. Yamanouchi, T. Matsuwaki, M. Nishihara (2012) Age-related resistance of skeletal muscle-derived progenitor cells to SPARC may explain a shift from myogenesis to adipogenesis. Aging 4: 40-48.
20.
Ock, S.A., G.J. Rho (2011) Effect of dimethyl sulfoxide (DMSO) on cryopreservation of porcine mesenchymal stem cells (pMSCs). Cell Transplant 20: 1231-1239.
21.
Oishi, K., H. Noguchi, H. Yukawa, T. Miyazaki, R. Kato, Y. Kitagawa, M. Ueda, S. Hayashi (2008) Cryopreservation of mouse adipose tissue-derived stem/progenitor cells. Cell Transplant 17: 35-41.
22.
Shah, F.S., J. Li, M. Dietrich, X. Wu, M.G. Hausmann, K.A. LeBlanc, J.W. Wade, J.M. Gimble (2014) Comparison of stromal/stem cells isolated from human omental and subcutaneous adipose depots: differentiation and immunophenotypic characterization. Cells Tissues Organs 200: 204-211.
23.
Thirumala, S., J.M. Gimble, R.V. Devireddy (2010a) Cryopreservation of stromal vascular fraction of adipose tissue in a serum-free freezing medium. J Tissue Eng Regen Med 4: 224-232.
24.
Thirumala, S., J.M. Gimble, R.V. Devireddy (2010b) 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 19: 513-522.
25.
Thirumala, S., X. Wu, J.M. Gimble, R.V. Devireddy (2010c) Evaluation of polyvinylpyrrolidone as a cryoprotectant for adipose tissue-derived adult stem cells. Tissue Eng Part C Methods 16: 783-792.
26.
Thirumala, S., S. Zvonic, E. Floyd, J.M. Gimble, R.V. Devireddy (2005) Effect of various freezing parameters on the immediate post-thaw membrane integrity of adipose tissue derived adult stem cells. Biotechnol Prog 21: 1511-1524.
27.
Ullah, M., S. Stich, M. Notter, J. Eucker, M. Sittinger, J. Ringe (2013) Transdifferentiation of mesenchymal stem cells-derived adipogenic-differentiated cells into osteogenic- or chondrogenic-differentiated cells proceeds via dedifferentiation and have a correlation with cell cycle arresting and driving genes. Differentiation 85: 78-90.
28.
Yong, K.W., B. Pingguan-Murphy, F. Xu, W.A. Abas, J.R. Choi, S.Z. Omar, M.A. Azmi, K.H. Chua, W.K. Wan Safwani (2015) Phenotypic and functional characterization of long-term cryopreserved human adipose-derived stem cells. Sci Rep 5: 9596.
29.
Yu, G., Z.E. Floyd, X. Wu, T. Hebert, Y.D. Halvorsen, B.M. Buehrer, J.M. Gimble (2011) Adipogenic differentiation of adipose-derived stem cells. Methods Mol Biol 702: 193-200.
30.
Yu, G., X. Wu, M.A. Dietrich, P. Polk, L.K. Scott, A.A. Ptitsyn, J.M. Gimble (2010) Yield and characterization of subcutaneous human adipose-derived stem cells by flow cytometric and adipogenic mRNA analyzes. Cytotherapy 12: 538-546.
31.
Zuk, P.A., M. Zhu, P. Ashjian, D.A. De Ugarte, J.I. Huang, H. Mizuno, Z.C. Alfonso, J.K. Fraser, P. Benhaim, M.H. Hedrick (2002) Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13: 4279-4295.
32.
Zuk, P.A., M. Zhu, H. Mizuno, J. Huang, J.W. Futrell, A.J. Katz, P. Benhaim, H.P. Lorenz, M.H. Hedrick (2001) Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 7: 211-228.
© 2016 S. Karger AG, Basel
2016
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.
