Vitrification Method Efficacy of Mesenchymal Stem Cells (MSCs) Derived from Wharton’s Jelly

Abstract

Abstract Cryopreservation is done to maintain the availability of stem cells for research and therapy. Vitrification is a cryopreservation method that has been used for frozen storage on embryonic stem cells (ESCs), induced pluripotent stem cell (iPS) and adult stem cells such as MSCs. However, the efficacy of these methods vary greatly depending on the use of cryoprotectants and the type of stem cells are stored, as well as in mesenchymal stem cells (MSCs) derived from umbilical cord matrix (Warthon's Jelly). This study aimed to determine the efficacy of cryopreservation by the vitrification method for suspension of MSCs from human Warthon's Jelly. Investigation was conducted in Stem Cell Laboratory, Center for Biomedical and Basic Technology of Health, NIHRD, Ministry of Health, Indonesia. MSC isolation and in vitro expansion were carried out prior to cryopreservation while evaluating their viability, population doubling (PD), population doubling time (PDT), immunophenotypes as well as karyotyping profile between control (non-vitrified) and vitrified group. In this study, we used two vitrification solutions: DAP-213 consisting of 10% 10M acetamide, 22% propylene glycol and 14.2% dimethyl sulfoxide (DMSO); and M20 composed of 20% DMSO dan ethylene glycol. Our study showed that viability, PD, and PDT of vitrified MSCs in DAP-213 and M20 were not different compared to control. Percentage of CD90 and CD73 expression for vitrified MSCs were lower than control (p 0,05). Pada penggunaan M20, PD dan PDT secara berurutan 2,48 dan 58,15 lebih rendah dibandingkan dengan kontrol dan DAP (p<0,05). Sedangkan ekspresi antigen CD90 dan CD73 MSC pasca vitrifikasi lebih rendah dibanding kontrol (p<0,05). Kesimpulan dari penelitian adalah metode vitrifikasi dapat digunakan untuk MSCs dari matriks tali pusat. Kata kunci : mesenchymal stem cell, vitrifikasi, Wharton’s Jelly, DAP-213, M20