title: Cryopreservation of Human Wharton's Jelly-derived Mesenchymal Stem Cells Following Controlled Rate Freezing Protocol Using Different Cryoprotectants; A Comparative Study.
abstract: OBJECTIVE
  To compare the effect of three different cryoprotectants on basic stem cell characteristics for the possibility of using well defined, dimethyl sulfoxide (DMSO) and serum free freezing solutions to cryopreserve human Wharton's jelly-derived mesenchymal stem cells (WJMSCs) following controlled rate freezing protocol.
  
  
  METHODS
  The mesenchymal stem cells isolated from human Wharton's jelly were cryopreserved using 10% DMSO, 10% polyvinylpyrrolidone (PVP) and a cocktail solution comprising of 0.05 M glucose, 0.05 M sucrose and 1.5 M ethylene glycol following controlled rate freezing protocol. We investigated the post-thaw cell viability, morphology, proliferation capacity, basic stem cell characteristics, in vitro differentiation potential and apoptosis-related gene expression profile before and after cryopreservation.
  
  
  RESULTS
  The cryoprotectant 10% DMSO has shown higher post-thaw cell viability of 81.2±0.58% whereas 10% PVP and cocktail solution have shown 62.87±0.35% and 72.2±0.23%, respectively at 0 h immediately thawing. The cell viability was further reduced in all the cryopreserved groups at 24 h later post-thaw culture. Further, the complete elimination of FBS in cryoprotectants has resulted in drastic reduction in cell viability. Cryopreservation did not alter the basic stem cell characteristics, plasticity and multipotency except proliferation rate. The expression of pro-apoptotic BAX and p53 genes were higher whilst p21 was lower in all the cryopreserved groups when compare to the control group of WJMSCs.
  
  
  CONCLUSIONS
  Although 10% DMSO has shown higher post-thaw cell viability compare to 10% PVP and cocktail solution, the present study indicates the feasibility of developing a well-defined DMSO free cryosolution which can improve storage and future broad range applications of WJMSCs in regenerative medicine without losing their basic stem cell characteristics.
authors.affiliation: Department of Vet OBS/Theriogenology and Biotechnology, Gyeongsang National University, Jinju, Korea.
authors.forename: Sharath Belame
authors.initials: SB
authors.lastname: Shivakumar
authors.affiliation: Department of Vet OBS/Theriogenology and Biotechnology, Gyeongsang National University, Jinju, Korea.
authors.forename: Dinesh
authors.initials: D
authors.lastname: Bharti
authors.affiliation: Department of Vet OBS/Theriogenology and Biotechnology, Gyeongsang National University, Jinju, Korea.
authors.forename: Si-Jung
authors.initials: SJ
authors.lastname: Jang
authors.affiliation: Department of Orthopaedic Surgery, School of Medicine, Gyeongsang National University, Jinju, Korea.
authors.forename: Sun-Chul
authors.initials: SC
authors.lastname: Hwang
authors.affiliation: Department of Obstetrics and Gynaecology, School of Medicine, Gyeongsang National University, Jinju, Korea.
authors.forename: Ji-Kwon
authors.initials: JK
authors.lastname: Park
authors.affiliation: Department of Obstetrics and Gynaecology, School of Medicine, Gyeongsang National University, Jinju, Korea.
authors.forename: Jeong-Kyu
authors.initials: JK
authors.lastname: Shin
authors.affiliation: Department of Oral and Maxillofacial Surgery, School of Medicine, Gyeongsang National University, Jinju, 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, Korea.
authors.forename: Bong-Wook
authors.initials: BW
authors.lastname: Park
authors.affiliation: Department of Vet OBS/Theriogenology and Biotechnology, Gyeongsang National University, Jinju, Korea ; Research Institute of Life Sciences, Gyeongsang National University, Jinju, Korea.
authors.forename: Gyu-Jin
authors.initials: GJ
authors.lastname: Rho
keywords: Apoptosis; Cocktail solution; Controlled rate freezing; Cryopreservation; Wharton’s jelly mesenchymal stem cells