Effect of mHCN2 gene modification on chronotropic relevant receptors in BMSCs co-cultured with atrial myocytes.

Currently, the mechanism of the chronotropic ability of stem cells modified to express the hyperpolarization-activated cyclic nucleotide-gated (HCN) gene remains to be elucidated. The present study assessed the effects of mouse (m)HCN2 gene modification on the expression of chronotropic relevant receptors, adrenergic receptor β1 (Adrb1) and cholinergic receptor muscarinic M2 (Chrm2), in bone marrow stromal cells (BMSCs) co-cultured with atrial myocytes. BMSCs were divided into the following four groups: i) BMSCs transfected with the mHCN2 gene and co-cultured with atrial myocytes for 48 h (TF + CO); ii) respective transfection (TF); iii) respective co-culture (CO); and iv) the control group without treatment (CTL). Green fluorescent protein (GFP) was observed in the BMSCs 48 h after transfection with pEGFP-C1-mHCN2. The expression of Adrb1 and Chrm2 was significantly increased in the TF and TF + CO groups, particularly the TF + CO group, compared with the CTL group (P<0.05). This suggests that BMSCs modified to express the mHCN2 gene possess autorhythmicity and chronotropic ability, particularly when co-cultured with atrial myocytes. The results of the present study provide novel information regarding the molecular basis of biological pacemakers' chronotropic ability.