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Br-DIF-1 Accelerates Dimethyl Sulphoxide-Induced Differentiation of P19CL6 Embryonic Carcinoma Cells Into Cardiomyocytes

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

Br J Pharmacol. 2012 Feb;165(4):870-9.

PMID: 21671902

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP111050727	DIF-1		111050-72-7	Price
AP118191303	Br-DIF-1		118191-30-3	Price

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