Objective: To transfect rat adipose tissue-derived adult stem cells (ADSCs) with the pacemaker gene of human hyperpolarization-activated cyclic nucleotide-gated ion channel 2 (hHCN2), and observe its feasibility of functioning as pacemaker cells.
Methods: First of all, healthy ADSCs were harvested and transfected with hHCN2 gene. Thereafter, we detected the expression of hHCN2 gene and protein by quantitative real-time PCR (qRT-PCR), Western blot analysis and immunofluorescence cytochemistry and measured the electrophysiological properties and recorded the inward currents using patch clamp techniques. In addition, we mixed and co-cultured the transfected ADSCs and rat cadiomyocytes to observe the influence on the spontaneous beating rate of cadiomyocytes.
Results: Compared with the control ADSCs, the transfected ADSCs expressed the higher level of hHCN2 gene and protein(P<0.05), and they could produce pacemaker current. When co-cultured with rat ventricular myocytes, they could accelerate the spontaneous beating rate of the ventricular myocytes(P<0.05).
Conclusion: After transfected with hHCN2 gene, ADSCs can work as pacemaker-like cells, which provides materials for the research of biological pacemakers.