Cardiovascular differentiation of monkey embryonic stem cell-derived vascular endothelial growth factor receptor-2-positive cells

Cardiovascular differentiation of monkey embryonic stem cell-derived vascular endothelial growth factor receptor-2-positive cells

Tomohide Takaya1, Teruhisa Kawamura2, Tatsuya Morimoto3, Koh Ono1, Takeshi Kimura1, Hirofumi Suemori4, Norio Nakatsuji4, Akira Shimatsu5, Koji Hasegawa5.

  1. Department of Cardiovascular Medicine, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
  2. Career-Path Promotion Unit for Young Life Scientists, Kyoto University, Kyoto, Japan.
  3. Clinical Pharmacology and Therapeutics, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka, Japan.
  4. Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan.
  5. Division of Translational Research, Kyoto Medical Center, National Hospital Organization, Kyoto, Japan.

The 6th China-Japan Cardiovascular Forum (Beijing, China), 2010/10/16 (Poster).

Abstract

Background: Human induced pluripotent stem (iPS) cells are potential sources of cardiovascular cells to be transplanted in patients with end-stage heart failure. The identification and purification of cardiovascular progenitor cells in primate ES/iPS cells in critical to apply human iPS cells for cardiovascular regenerative therapy in the clinical setting. Cynomolgus monkey Macaca fascicularis embryonic stem (ES) cells have similar properties to human ES/iPS cells and can be used for characterization of primate stem cells.

Methods and Results: Monkey ES cells were maintained on a mouse embryonic fibroblast feeder layer. Throughout this study, knockout serum replacement was used instead of animal-derived serum. Monkey ES cells were differentiated by forming embryoid bodies (EBs). Expression of a mesodermal marker, vascular endothelial growth factor receptor-2 (VEGFR-2), and an undifferentiation marker, tumor-related antigen-1-81 (TRA-1-81), were analyzed by flowcytometer. Their expression patterns of monkey ES cells were similar to those of human ES cells, but not those of mouse ES cells. The monkey VEGFR-2+/TRA-1-81- cells were collected from EBs at day 7-9, then cocultured with neonatal rat cardiomyocytes in the serum-free condition. After 7 days of coculture, the monkey VEGFR-2+/TRA-1-81- cells expressed markers of cardiac muscle cells (GATA4, Nkx2.5, ANF, and β-MHC) and those of vascular cells (CD31, VE-cadherin, and smooth muscle actin). However, these cells did not express ectoderm (PAX6) and endoderm (SOX17 and α-fetoprotein) markers.

Conclusions: VEGFR-2+/TRA-1-81- cells derived from monkey ES cells cocultured with rat cardiomyocytes in the serum-free condition are able to differentiate specifically into cardiovascular cells. This system will be useful to determine and purify cardiovascular progenitor cells derived from human iPS cells in regenerative therapy for end-staged heart failure.