DMSO- and Serum-Free Cryopreservation of Wharton's Jelly Tissue Isolated From Human Umbilical Cord.
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.
['Shivakumar|Sharath Belame|SB|', 'Bharti|Dinesh|D|', 'Subbarao|Raghavendra Baregundi|RB|', 'Jang|Si-Jung|SJ|', 'Park|Ji-Sung|JS|', 'Ullah|Imran|I|', 'Park|Ji-Kwon|JK|', 'Byun|June-Ho|JH|', 'Park|Bong-Wook|BW|', 'Rho|Gyu-Jin|GJ|']
D017209:Apoptosis D015153:Blotting, Western D002454:Cell Differentiation D049109:Cell Proliferation D002478:Cells, Cultured D015925:Cryopreservation D003451:Cryoprotective Agents D004121:Dimethyl Sulfoxide D005260:Female D006801:Humans D008297:Male D059630:Mesenchymal Stem Cells D012333:RNA, Messenger D060888:Real-Time Polymerase Chain Reaction D020133:Reverse Transcriptase Polymerase Chain Reaction D014470:Umbilical Cord D059631:Wharton Jelly
WHARTON'S JELLY TISSUE FREEZING CRYOPROTECTANTS COCKTAIL SOLUTION CELL SURVIVAL RATE WHARTON'S JELLY MESENCHYMAL STEM CELLS