Phenotypic and Functional Characterization of Long-Term Cryopreserved Human Adipose-derived Stem Cells

Yong, K.W. and Pingguan-Murphy, Belinda and Xu, F. and Wan Abas, Wan Abu Bakar and Choi, J.R. and Omar, Siti Zawiah and Noor Azmi, Mat Adenan and Chua, K.H. and Wan Kamarul Zaman, Wan Safwani (2015) Phenotypic and Functional Characterization of Long-Term Cryopreserved Human Adipose-derived Stem Cells. Scientific Reports, 5 (1). p. 9596. ISSN 2045-2322, DOI

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Cryopreservation represents an effective technique to maintain the functional properties of human adipose-derived stem cells (ASCs) and allows pooling of cells via long-term storage for clinical applications, e.g., cell-based therapies. It is crucial to reduce freezing injury during the cryopreservation process by loading the ASCs with the optimum concentration of suitable cryoprotective agents (CPAs). In this study, human ASCs were preserved for 3 months in different combinations of CPAs, including 1) 0.25 M trehalose; 2) 5% dimethylsulfoxide (DMSO); 3) 10% DMSO; 4) 5% DMSO + 20% fetal bovine serum (FBS); 5) 10% DMSO + 20% FBS; 6) 10% DMSO + 90% FBS. Interestingly, even with a reduction of DMSO to 5% and without FBS, cryopreserved ASCs maintained high cell viability comparable with standard cryomedium (10% DMSO + 90% FBS), with normal cell phenotype and proliferation rate. Cryopreserved ASCs also maintained their differentiation capability (e.g., to adipocytes, osteocytes and chondrocytes) and showed an enhanced expression level of stemness markers (e.g., NANOG, OCT-4, SOX-2 and REX-1). Our findings suggest that 5% DMSO without FBS may be an ideal CPA for an efficient long-term cryopreservation of human ASCs. These results aid in establishing standardized xeno-free long-term cryopreservation of human ASCs for clinical applications.

Item Type: Article
Funders: University of Malaya from the Ministry of Education Malaysia: High Impact Research Grant (UM.C/HIR/MOHE/ENG/44) and Postgraduate Research Grant (PPP) (PG007-2014A), National Natural Science Foundation of China (11372243), International Science & Technology Cooperation Program of China (2013DFG02930), Major International Joint Research Program of China (11120101002), China Young 1000-Talent Program, Program for New Century Excellent Talents in University
Uncontrolled Keywords: Adipogenesis; Adipose Tissue; Adult; Adult Stem Cells; Cell Culture Techniques; Cell Differentiation; Cell Proliferation; Cell Self Renewal; Cell Separation; Cell Survival; Chondrogenesis; Cryopreservation; Female; Humans; Immunophenotyping; Osteogenesis; Phenotype; Time Factors; Young Adult
Subjects: R Medicine
Divisions: Faculty of Medicine
Depositing User: Ms. Juhaida Abd Rahim
Date Deposited: 21 Sep 2018 03:12
Last Modified: 16 Dec 2019 03:36

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