SULMAZ KAMALİFAR, NEGAR AZARPİRA, LADAN SADEGHİ, SADEGH GHORBANİ-DALİNİ, SEİDEH MASOOMEH NEKOEİ, MAHDOKHT H AGHDAİE, ELAHEH ESFANDİARİ, MOHAMAD REZA AZARPİRA
Experimental and Clinical Transplantation - 2020;18(4):505-511
Objectives: Wharton jelly mesenchymal stem cells are good candidates for application in different aspects of regenerative medicine, and their long-time banking is important. In this study, the effects of trehalose, ascorbic acid, and Y-27632 on proliferation and survival rate of these cells after cryopreservation were investigated. Materials and Methods: Mesenchymal stem cells were isolated from human umbilical cord Wharton jelly and frozen using a slow-rate cooling process. Different concentrations of trehalose (35, 75, and 125 mM), ascorbic acid (0.06, 0.125, 0.25, and 0.5 mM), and Y-27632 (10 μM) were used to treat culture medium and/or to supplement freezing medium. Assessment of cell viability after thawing was performed using Trypan blue staining, and MTT assay was performed to measure the cell proliferation rate. Results: We observed significantly increased postthaw viability, increased cell proliferation, and decreased doubling time of cells when 75 mM trehalose, 0.25 and 0.5 mM ascorbic acid, and 10 mM Y-27632 were used. In addition, increased viability, proliferation, and attachment were observed after 24 hours of pre - treatment with these cryoprotective agents and when they were added to conventional freezing medium. Conclusions: The use of different cryoprotective agents in culture and freezing media could be useful for longterm storage of Wharton jelly mesenchymal stem cells.
