BACKGROUND: Adipose-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 degree 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.
