The Effect of Immunosuppression Agents on Stem and Endothelial Cells’ Proliferation in Vitro

The control of immune-mediated transplant rejection by immunosuppressive agents might affect other human cells’ proliferation and function. This study aims to evaluate the effect of three immunosuppressants on the proliferation of human adipose-derived adult stroma (ADAS) and human umbilical vein endothelial cells (HUVEC). These cells were used as a culture model to study the minimum inhibitory dose of commonly used immunosuppressants. Cells were cultured and seeded into 96-well plates at a density of 2,500/well then treated either with one of two calcineurin inhibitors, namely FK506 (Tacrolimus) and Cyclosporin A (CsA), or with the IMP dehydrogenase inhibitor, Mycophenolic acid (MPA). After 5 days, the medium was removed and plates were stored at −70°C until analyzed with the CyQUANT cell proliferation assay. The minimum inhibitory dose on HUVEC proliferation was 1000 ng/ml for CsA, 500 ng/ml for MPA, and 10,000 ng/ml for FK506. The minimum inhibitory dose on ADAS proliferation was 5 ng/ml for MPA, 100000 ng/ml for FK506 and 100000 ng/ml for CsA. Our results suggest that these immunosuppressants inhibit ADAS and HUVEC proliferation in a dose-dependent manner in vitro. The FK506 and CsA minimum inhibitory concentrations on both HUVEC and ADAS were several orders of magnitude greater than those required for T cell inhibition and, therefore, regular dosages are not expected to impair HUVEC or ADAS proliferation. However, the minimum inhibitory concentration of MPA on ADAS was lower than that observed in vitro for lymphocyte function and, therefore, could be of concern. Experimental article (J Int Dent Med Res 2015; 8: (3), pp. 116-122)