Effect of Fasudil Hydrochloride on the post-thaw Viability of Cryopreserved Porcine Adipose-Derived Stem Cells

BACKGROUND: A dipose-derived stem cells (ADSCs) are of interest for regenerative medicine as they are isolated easily and can differentiate into multiple cell lineages. Recently, it was reported that a Rho-associated kinase (ROCK) inhibitor Y-27632 could enhance the post-thaw viability and physiological function of cryopreserved BMSC. OBJECTIVE: The present study is to investigate whether Fasudil hydrochloride (FH, a selective Rho-kinase inhibitor like Y-27632) can exert a similar beneficial effect on the post-thaw viability of cryopreserved ADSCs. METHODS: ADSCs were allotted in three cryopreservation solutions [I:10% (v/v) (DMSO), 30% (v/v) FBS (as a negative control);II:10μM FH, 10% (v/v) DMSO, 30% (v/v) FBS;III:10 μM Y-27632, 10% (v/v) DMSO, 30% (v/v) fetal bovine serum , All groups were frozen using a rapid freezing method and stored at -196° C in liquid nitrogen for 30 days. After thawing and being cultured 24h, viability of ADSCs were detected by MTT assay. RESULT: The MTT assay showed significant differences in the proportion of adherent viable cells over the concentration of Y-27632 and FH, Additionally, FH did not induce morphological changes in the frozen-thawed ADSCs like Y-27632 did. CONCLUSIONS: FH might represent a promising cryopreservation solution in enhancing the post-thaw viability and physiological function of cryopreserved ADSCs and did not chang the cell appearance.