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

Research output: Contribution to journalArticle

21 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

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Endothelial cells
Embryonic Stem Cells
Stem cells
Endothelial Cells
Vascular Endothelial Growth Factor A
rho-Associated Kinases
Therapeutic Uses
Protein Kinase Inhibitors
Phosphatidylinositol 3-Kinases
Chemical activation

ASJC Scopus subject areas

  • Hematology
  • Biochemistry
  • Cell Biology
  • Immunology

Cite this

ROCK suppression promotes differentiation and expansion of endothelial cells from embryonic stem cell-derived Flk1+ mesodermal precursor cells. / Joo, Hyung Joon; Choi, Dong Kyu; Lim, Joon Seo; Park, Jin Sung; Lee, Seung Hun; Song, Sukhyun; Shin, Jennifer H.; Lim, Do-Sun; Kim, Injune; Hwang, Ki Chul; Koh, Gou Young.

In: Blood, Vol. 120, No. 13, 27.09.2012, p. 2733-2744.

Research output: Contribution to journalArticle

Joo, Hyung Joon ; Choi, Dong Kyu ; Lim, Joon Seo ; Park, Jin Sung ; Lee, Seung Hun ; Song, Sukhyun ; Shin, Jennifer H. ; Lim, Do-Sun ; Kim, Injune ; Hwang, Ki Chul ; Koh, Gou Young. / ROCK suppression promotes differentiation and expansion of endothelial cells from embryonic stem cell-derived Flk1+ mesodermal precursor cells. In: Blood. 2012 ; Vol. 120, No. 13. pp. 2733-2744.
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AU - Lee, Seung Hun

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AU - Shin, Jennifer H.

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