Myogenic oligodeoxynucleotide, myoDN, induces myocardial differentiation of murine iPS cells
Mina Ishioka, Yuma Nihashi, Hiroshi Kagami, Takeshi Shimosato, Tomohide Takaya.
Faculty of Agriculture, Shinshu University, Nagano, Japan.
2018 International Symposium on Animal Science and Technology (Ina, Japan), 2018/08/10 (Poster).
Abstract
Summary: We recently identified the 18 nt-oligodeoxynucleotide, named myoDN, which intensively induces myogenic differentiation of skeletal muscle myoblasts. In this study, we investigated the effects of myoDN on differentiation of murine induced pluripotent stem (iPS) cells.
Materials & Methods: iPS cell line 20D17 was generated by introducing Oct3/4, Sox2, Klf4, and c-Myc into murine embryonic fibroblasts (MEF) in which a GFP gene is inserted in Nanog locus (Okita, Nature, 2007). Undifferentiated 20D17 cells were maintained on a MEF feeder layer. To induce differentiation, 3×104 of 20D17 cells were seeded on gelatin-coated dishes (defined as day 0) and cultured in differentiation medium (DMEM, 10% fetal bovine serum, 5% horse serum, non-essential amino acids, and β-mercaptoethanol) (Mizuno, FASEB J, 2010). The cells were stimulated with 10 uM myoDN or its vehicle as a negative control during days 5-9.
Results & Discussion: In control group, some beating colonies of the spontaneously-differentiated cardiomyocytes were observed at day 11. Diameters of the clusters were around 300 um, and the beating rate was a maximum of 150 times/min by day 15. While in myoDN-treated group, numerous beating clusters were observed all over the dishes. Diameters and beating rates of the clusters in myoDN group reached at least 600 um and over 200 times/min, respectively. These results indicate that myoDN is able to induce myocardial differentiation of pluripotent stem cells. myoDN may be a useful tool for regenerative medicine in the heart.