The Rho-associated kinase inhibitor fasudil can replace Y-27632 for use in human pluripotent stem cell research
Fig 2
Fasudil enhances the cell growth of hPSCs.
A: Growth of hES3 cells for 7 days according to fasudil concentration. The growth of hES3 cells at fasudil concentrations of 1, 5, and 10 μM was concentration-independent, and significantly faster than the untreated control (*: p < 0.05, 3 biological replicates). B: ALP staining of hES3 and hiPS2 cells cultured with Y-27632 or fasudil. Treatment with fasudil or Y-27632 increased the size of PSC colonies. Scale bar = 10 mm. C, D: Fold increase of initially seeded hESCs (hES1, 2, and 3) and hiPSCs (hiPS1 and 2) using commercial reagent C or lab-made reagent D. Growth of all hPSC lines except hES1 increased in response to fasudil or Y-27632 (*: p < 0.05, 3 biological replicates). E: Increase of hES3 cell numbers with time. After three passages, treatment with fasudil and Y-27632 significantly increased cell numbers compared with the untreated group (*: p < 0.05, 3 technical replicates). hPSC: human pluripotent stem cell, hES: human embryonic stem cell, ALP: Alkaline Phosphatase, hiPS: human induced pluripotent stem cell.