Sinoatrial node cells phenotype expression is induced by direct bone marrow stem cells contact with sinoatrial node cells
Guan, S.-b.; Ma, A.-q.; Jiang, W.-h.
Zhonghua Xin Xue Guan Bing Za Zhi 37(1): 73-76
2009
ISSN/ISBN: 0253-3758 PMID: 19671359 Document Number: 632817
To observe the differentiation of bone mesenchymal stem cells (BMSCs) co-cultured with purified sinoatrial node cells (SNC) of neonate rats. SNC from neonatal SD rat were cultured and purified with differential attachment method and labeled with BrdU. Rat BMSCs were isolated by a Percoll's gradient solution and cultured in DMEM. After 2 passages, these BMSCs were transfected with pEGFP-N1 by Lipofectamine and labeled with GFP. EGFP-BMSC were co-cultured with SNC in a rate of 1:5 for 1 week. EGFP-BMSC cultured in SNC culture medium served as controls. SNC marker hyperpolarization activated cyclic nucleotide gated cation channel 4 (HCN4) and connexin 45 (Cx45) expressions were determined by immunofluorescence staining. Positive immunofluorescence staining against HCN4 and Cx45 were detected in EGFP-BMSC co-cultured with SNC but not in EGFP-BMSC cultured in SNC culture medium. Direct cell-to-cell contact between BMSCs and SNC cells may induce BMSCs differentiation into sinus node-like cells.