Br-DIF-1 accelerates dimethyl sulphoxide-induced differentiation of P19CL6 embryonic carcinoma cells into cardiomyocytes

K. Seya, K. Kanemaru, M. Matsuki, K. Hongo, H. Kitahara, H. Kikuchi, Y. Oshima, Y. Kubohara, K. Okumura, S. Motomura, K. I. Furukawa

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9 Citations (Scopus)

Abstract

BACKGROUND AND PURPOSE Stem cell transplantation therapy is a promising option for treatment of severe ischaemic heart disease. Dimethyl sulphoxide (DMSO) differentiates P19CL6 embryonic carcinoma cells into cardiomyocyte-like cells, but with low differentiation capacity. To improve the degree of this differentiation, we have assessed several derivatives of the differentiation-inducing factor-1 (DIF-1), originally found in the cellular slime mould Dictyostelium discoideum, on P19CL6 cells. EXPERIMENTAL APPROACH P19CL6 cells were cultured with each derivative and 1% DMSO for up to 16 days. Differentiation was assessed by measuring the number of beating and non-beating aggregates, and the expression of genes relevant to cardiac tissue. The mechanism of action was investigated using a T-type Ca 2+ channel blocker. KEY RESULTS Of all the DIF-1 derivatives tested only Br-DIF-1 showed any effects on cardiomyocyte differentiation. In the presence of 1% DMSO, Br-DIF-1 (0.3-3 μM) significantly and dose-dependently increased the number of spontaneously beating aggregates compared with 1% DMSO alone, by day 16. Expression of mRNA for T-type calcium channels was significantly increased by Br-DIF-1 + 1% DMSO compared with 1% DMSO alone. Mibefradil (a T-type Ca 2+ channel blocker; 100 nM) and a small interfering RNA for the T-type Ca 2+ channel both significantly decreased the beating rate of aggregates induced by Br-DIF-1 + 1% DMSO. CONCLUSIONS AND IMPLICATIONS Br-DIF-1 accelerated the differentiation, induced by 1% DMSO, of P19CL6 cells into spontaneously beating cardiomyocyte-like cells, partly by enhancing the expression of the T-type Ca 2+ channel gene.

Original languageEnglish
Pages (from-to)870-879
Number of pages10
JournalBritish Journal of Pharmacology
Volume165
Issue number4
DOIs
Publication statusPublished - 2012 Feb
Externally publishedYes

Keywords

  • beating
  • Br-DIF-1
  • cardiomyocyte differentiation
  • dimethyl sulphoxide
  • mibefradil
  • mouse
  • P19CL6
  • siRNA
  • T-type Ca channel

ASJC Scopus subject areas

  • Pharmacology

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