Induction of cardiac differentiation of pluripotent stem cells by myogenetic oligodeoxynucleotide

Induction of cardiac differentiation of pluripotent stem cells by myogenetic oligodeoxynucleotide

Mina Ishioka1, Yuma Nihashi2, Koji Umezawa1,2,3, Hiroshi Kagami1,2, Takeshi Shimosato1,2,3, Tomohide Takaya1,2,3.

  1. Graduate School of Science and Technology, Shinshu University.
  2. Graduate School of Medicine, Science and Technology, Shinshu University.
  3. Institute for Biomedical Sciences, Shinshu University.

日本核酸医薬学会第5回年会 (大阪), 2019/07/10 (口演).

Abstract

【Introduction】We recently identified a telomeric 18 nt-oligodeoxynucleotide, myoDN, which intensively induces differentiation of skeletal muscle myoblasts. In this study, we investigated the effect of myoDN on differentiation of pluripotent stem cells.

【Methods】We utilized murine embryonic stem cell (mESC) line, hCGp7, in which GFP was knocked into the locus of a cardiac specific gene Nkx2-5. hCGp7 was seeded on gelatin-coated dishes and induced spontaneous differentiation in DMEM/10% FBS/5% HS. The cells were treated with 10 uM myoDN on and after day 3.

【Results】In the hCGp7 with myoDN from day 5, numerous GFP+ beating colonies appeared on day8, indicating that myoDN induces cardiac differentiation of mESCs. Next, incidence ratio of beating colonies were measured when myoDN stimulation was started on day 3-7. In myoDN(-) group, beating ratio was reached 100% on day 10. In the hCGp7 with myoDN from day 4 or 5, all samples showed beating colonies on day 7 or 8. While, the hCGp7 with myoDN from day 3 never generated beating cells by day 11, resulting complete inhibition of cardiac differentiation. These data suggest myoDN switches cardiac cell fate depending on differentiation stages of mESCs. This study presented that simple treatment of myoDN easily induces cardiomyocytes from pluripotent stem cells, which will be useful for regenerative medicine or drug screening.