Application of the oligodeoxynucleotide from lactic acid bacteria that promotes myogenic differentiation

Application of the oligodeoxynucleotide from lactic acid bacteria that promotes myogenic differentiation

Sayaka Shinji, Yuma Nihashi, Koji Umezawa, Takeshi Shimosato, Tomohide Takaya.

Faculty of Agriculture, Shinshu University, Nagano, Japan.

2017 Japan-Thailand International Symposium on Animal Biotechnology (Ina, Japan), 2017/07/05 (Poster).

Abstract

Summary: We recently found that the oligodeoxynucleotide, involved in the genome of Lactobacillus rhamnosus GG, intensively promotes myogenic differentiation of skeletal muscle myoblasts. This oligonucleotide named "myoDN" induced differentiation of myoblasts but did not have any effects on fibroblasts. It is anticipated that such specific action of myoDN is useful to treat rhabdomyosarcoma (RMS; a myogenic tumor) and to generate skeletal muscle cells from pluripotent stem cells. In this study, we investigated whether myoDN acts on RMS cells and induced pluripotent stem cells (iPSCs) in vitro.

Materials & Methods: Human RMS cell line, KYM1, was cultured with myoDN. To evaluate their growth, the number of the cells and expression levels of cell cycle genes were quantified.

Myogenic differentiation of murine iPSCs were induced by forming embryoid bodies (EBs) in suspension culture. EBs were then plated on collagen-coated dishes and treated with myoDN. Myogenic gene expression of the EBs was examined by qPCR.

Results & Discussion: The number of KYM1 cells was significantly decreased after 48 h of myoDN treatment. qPCR results suggested that myoDN suppresses proliferation of KYM1 through promoting myogenic differentiation. Intriguingly, the effects of myoDN were different among RMS cell lines, which may be important to understand the mechanism of myoDN function.

myoDN also promoted skeletal muscle differentiation of iPSCs. myoDN treatment enhanced the expression of marker genes for terminally differentiated myocytes. myoDN is possibly helpful to generate mature skeletal muscle cells from patient-derived iPSCs for regenerative medicine.