title: DMSO- and Serum-Free Cryopreservation of Wharton's Jelly Tissue Isolated From Human Umbilical Cord.
abstract: The facile nature of mesenchymal stem cell (MSC) acquisition in relatively large numbers has made Wharton's jelly (WJ) tissue an alternative source of MSCs for regenerative medicine. However, freezing of such tissue using dimethyl sulfoxide (DMSO) for future use impedes its clinical utility. In this study, we compared the effect of two different cryoprotectants (DMSO and cocktail solution) on post-thaw cell behavior upon freezing of WJ tissue following two different freezing protocols (Conventional [-1°C/min] and programmed). The programmed method showed higher cell survival rate compared to conventional method of freezing. Further, cocktail solution showed better cryoprotection than DMSO. Post-thaw growth characteristics and stem cell behavior of Wharton's jelly mesenchymal stem cells (WJMSCs) from WJ tissue cryopreserved with a cocktail solution in conjunction with programmed method (Prog-Cock) were comparable with WJMSCs from fresh WJ tissue. They preserved their expression of surface markers, pluripotent factors, and successfully differentiated in vitro into osteocytes, adipocytes, chondrocytes, and hepatocytes. They also produced lesser annexin-V-positive cells compared to cells from WJ tissue stored using cocktail solution in conjunction with the conventional method (Conv-Cock). Real-time PCR and Western blot analysis of post-thaw WJMSCs from Conv-Cock group showed significantly increased expression of pro-apoptotic factors (BAX, p53, and p21) and reduced expression of anti-apoptotic factor (BCL2) compared to WJMSCs from the fresh and Prog-Cock group. Therefore, we conclude that freezing of fresh WJ tissue using cocktail solution in conjunction with programmed freezing method allows for an efficient WJ tissue banking for future MSC-based regenerative therapies. J. Cell. Biochem. 117: 2397-2412, 2016. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501, Jinju-daero, Jinju, 660-701, Republic of Korea.
authors.forename: Sharath Belame
authors.initials: SB
authors.lastname: Shivakumar
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501, Jinju-daero, Jinju, 660-701, Republic of Korea.
authors.forename: Dinesh
authors.initials: D
authors.lastname: Bharti
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501, Jinju-daero, Jinju, 660-701, Republic of Korea.
authors.forename: Raghavendra Baregundi
authors.initials: RB
authors.lastname: Subbarao
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501, Jinju-daero, Jinju, 660-701, Republic of Korea.
authors.forename: Si-Jung
authors.initials: SJ
authors.lastname: Jang
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501, Jinju-daero, Jinju, 660-701, Republic of Korea.
authors.forename: Ji-Sung
authors.initials: JS
authors.lastname: Park
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501, Jinju-daero, Jinju, 660-701, Republic of Korea.
authors.forename: Imran
authors.initials: I
authors.lastname: Ullah
authors.affiliation: Department of Obstetrics and Gynecology, School of Medicine, Gyeongsang National University, Jinju, 660-702, Republic of Korea.
authors.forename: Ji-Kwon
authors.initials: JK
authors.lastname: Park
authors.affiliation: Department of Oral and Maxillofacial Surgery, School of Medicine, Gyeongsang National University, Jinju, 660-702, Republic of Korea.
authors.forename: June-Ho
authors.initials: JH
authors.lastname: Byun
authors.affiliation: Department of Oral and Maxillofacial Surgery, School of Medicine, Gyeongsang National University, Jinju, 660-702, Republic of Korea.
authors.forename: Bong-Wook
authors.initials: BW
authors.lastname: Park
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, 501, Jinju-daero, Jinju, 660-701, Republic of Korea.|Research Institute of Life Sciences, Gyeongsang National University, 501, Jinju-daero, Jinju, 660-701, Republic of Korea.
authors.forename: Gyu-Jin
authors.initials: GJ
authors.lastname: Rho
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D005260:Female; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014470:Umbilical Cord / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D059631:Wharton Jelly / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism
chemical_list: D003451:Cryoprotective Agents; D012333:RNA, Messenger; D004121:Dimethyl Sulfoxide
keywords: CELL SURVIVAL RATE; COCKTAIL SOLUTION; CRYOPROTECTANTS; FREEZING; WHARTON'S JELLY MESENCHYMAL STEM CELLS; WHARTON'S JELLY TISSUE