Abstract
Efficient derivation of endothelial cells and their progenitors from human pluripotent stem cells (hPSCs) can facilitate studies of human vascular development, disease modeling, drug discovery, and cell-based therapy. Here we provide a detailed protocol for directing hPSCs to functional endothelial cells and their progenitors in a completely defined, growth factor- and serum-free system by temporal modulation of Wnt/β-catenin signaling via small molecules. We demonstrate a 10-day, two-stage process that recapitulates endothelial cell development, in which hPSCs first differentiate to endothelial progenitors that then generate functional endothelial cells and smooth muscle cells. Methods to characterize endothelial cell identity and function are also described.
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References
Kaupisch A, Kennedy L, Stelmanis V et al (2012) Derivation of vascular endothelial cells from human embryonic stem cells under GMP-compliant conditions: towards clinical studies in ischaemic disease. J Cardiovasc Transl Res 5:605–617
Levenberg S, Golub JS, Amit M et al (2002) Endothelial cells derived from human embryonic stem cells. Proc Natl Acad Sci U S A 99:4391–4396
Van der Meer AD, Orlova VV, ten Dijke P et al (2013) Three-dimensional co-cultures of human endothelial cells and embryonic stem cell-derived pericytes inside a microfluidic device. Lab Chip 13:3562–3568
Wilson HK, Canfield SG, Shusta EV, Palecek SP (2014) Concise review: tissue-specific microvascular endothelial cells derived from human pluripotent stem cells. Stem Cells 32:3037–3045
Ashton RS, Keung AJ, Peltier J, Schaffer DV (2011) Progress and prospects for stem cell engineering. Annu Rev Chem Biomol Eng 2:479–502
Lian X, Selekman J, Bao X et al (2013) A small molecule inhibitor of SRC family kinases promotes simple epithelial differentiation of human pluripotent stem cells. PLoS One 8:e60016
Lian X, Bao X, Zilberter M et al (2015) Chemically defined, albumin-free human cardiomyocyte generation. Nat Methods 12:595–596
Murry CE, Keller G (2008) Differentiation of embryonic stem cells to clinically relevant populations: lessons from embryonic development. Cell 132:661–680
Choi K-D, Yu J, Smuga-Otto K et al (2009) Hematopoietic and endothelial differentiation of human induced pluripotent stem cells. Stem Cells 27:559–567
Vodyanik MA, Thomson JA, Slukvin II (2006) Leukosialin (CD43) defines hematopoietic progenitors in human embryonic stem cell differentiation cultures. Blood 108:2095–2105
James D, Nam H, Seandel M et al (2010) Expansion and maintenance of human embryonic stem cell-derived endothelial cells by TGFbeta inhibition is Id1 dependent. Nat Biotechnol 28:161–166
Rufaihah AJ, Huang NF, Jamé S et al (2011) Endothelial cells derived from human iPSCS increase capillary density and improve perfusion in a mouse model of peripheral arterial disease. Arterioscler Thromb Vasc Biol 31:e72–e79
Goldman O, Feraud O, Boyer-Di Ponio J et al (2009) A boost of BMP4 accelerates the commitment of human embryonic stem cells to the endothelial lineage. Stem Cells 27:1750–1759
Sahara M, Hansson EM, Wernet O et al (2014) Manipulation of a VEGF-Notch signaling circuit drives formation of functional vascular endothelial progenitors from human pluripotent stem cells. Cell Res 24:820–841
White MP, Rufaihah AJ, Liu L et al (2013) Limited gene expression variation in human embryonic stem cell and induced pluripotent stem cell-derived endothelial cells. Stem Cells 31:92–103
Tatsumi R, Suzuki Y, Sumi T et al (2011) Simple and highly efficient method for production of endothelial cells from human embryonic stem cells. Cell Transplant 20:1423–1430
Wang ZZ, Au P, Chen T et al (2007) Endothelial cells derived from human embryonic stem cells form durable blood vessels in vivo. Nat Biotechnol 25:317–318
Bao X, Lian X, Dunn KK et al (2015) Chemically-defined albumin-free differentiation of human pluripotent stem cells to endothelial progenitor cells. Stem Cell Res 15:122–129
Lian X, Bao X, Al-Ahmad A et al (2014) Efficient differentiation of human pluripotent stem cells to endothelial progenitors via small-molecule activation of WNT signaling. Stem Cell Reports 3:804–816
Lian X, Zhang J, Azarin SM et al (2013) Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt/β-catenin signaling under fully defined conditions. Nat Protoc 8:162–175
Acknowledgments
This study was supported by NIH grant R01 EB007534 and NSF grant EFRI 0735903.
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Bao, X., Lian, X., Palecek, S.P. (2016). Directed Endothelial Progenitor Differentiation from Human Pluripotent Stem Cells Via Wnt Activation Under Defined Conditions. In: Barrett, Q., Lum, L. (eds) Wnt Signaling. Methods in Molecular Biology, vol 1481. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6393-5_17
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DOI: https://doi.org/10.1007/978-1-4939-6393-5_17
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6391-1
Online ISBN: 978-1-4939-6393-5
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