ROCK suppression promotes differentiation and expansion of endothelial cells from embryonic stem cell-derived Flk1+ mesodermal precursor cells

Hyung Joon Joo, Dong Kyu Choi, Joon Seo Lim, Jin Sung Park, Seung Hun Lee, Sukhyun Song, Jennifer H. Shin, Do-Sun Lim, Injune Kim, Ki Chul Hwang, Gou Young Koh

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25 Citations (Scopus)

Abstract

Successful differentiation and expansion of endothelial cells (ECs) from embryonic stem cell (ESC)-derived Flk1+ mesodermal precursor cells (MPCs) requires supplementation of vascular endothelial growth factor-A (VEGF-A). While analyzing VEGF-A/VEGFR2 downstream signaling pathway that underlies the VEGF-Ainduced differentiation and expansion of ECs, we fortuitously found that Rho-associated protein kinase (ROCK) inhibitor Y27632 profoundly promoted the differentiation and expansion of ECs from Flk1 + MPCs while reducing the differentiation and expansion of mural cells. The ROCK suppression-induced expansion of ECs appears to have resulted from promotion of proliferation of ECs via activation of PI3-kinase-Akt signaling. The ECs obtained by the combination of ROCK suppression and VEGF-A supplementation faithfully expressed most pan-EC surface makers, and phenotypic analyses revealed that they were differentiated toward arterial EC. Further incubation of the ICAM2+ ECs with Y27632 and VEGF-A for 2 days promoted expansion of ECs by 6.5-fold compared with those incubated with only VEGF-A. Importantly, the ROCK suppression-induced ECs displayed neovasculogenic abilities in vitro and in vivo. Thus, supplementation of ROCK inhibitor Y27632 along with VEGF-A in 2D Matrigel culture system provides a simple, efficient, and versatile method for obtaining ample amount of ESC-derived ECs at high purity suitable for use in therapeutic neovascularization.

Original languageEnglish
Pages (from-to)2733-2744
Number of pages12
JournalBlood
Volume120
Issue number13
DOIs
Publication statusPublished - 2012 Sep 27

ASJC Scopus subject areas

  • Hematology
  • Biochemistry
  • Cell Biology
  • Immunology

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    Joo, H. J., Choi, D. K., Lim, J. S., Park, J. S., Lee, S. H., Song, S., Shin, J. H., Lim, D-S., Kim, I., Hwang, K. C., & Koh, G. Y. (2012). ROCK suppression promotes differentiation and expansion of endothelial cells from embryonic stem cell-derived Flk1+ mesodermal precursor cells. Blood, 120(13), 2733-2744. https://doi.org/10.1182/blood-2012-04-421610