title: Human induced pluripotent stem cell-derived endothelial cells exhibit functional heterogeneity.
abstract: Human induced pluripotent stem cell-derived endothelial cells (hiPSC-ECs) are promising for treatment of vascular diseases. However, hiPSC-ECs purified based on CD31 expression are comprised of arterial, venous, and lymphatic subtypes. It is unclear whether hiPSC-ECs are heterogeneous in nature, and whether there may be functional benefits of enriching for specific subtypes. Therefore, we sought to characterize the hiPSC-ECs and enrich for each subtype, and demonstrate whether such enrichment would have functional significance. The hiPSC-ECs were generated from differentiation of hiPSCs using vascular endothelial growth factor (VEGF)-A and bone morphogenetic protein-4. The hiPSC-ECs were purified based on positive expression of CD31. Subsequently, we sought to enrich for each subtype. Arterial hiPSC-ECs were induced using higher concentrations of VEGF-A and 8-bromoadenosine-3':5'-cyclic monophosphate in the media, whereas lower concentrations of VEGF-A favored venous subtype. VEGF-C and angiopoietin-1 promoted the expression of lymphatic phenotype. Upon FACS purification based on CD31+ expression, the hiPSC-EC population was observed to display typical endothelial surface markers and functions. However, the hiPSC-EC population was heterogeneous in that they displayed arterial, venous, and to a lesser degree, lymphatic lineage markers. Upon comparing vascular formation in matrigel plugs in vivo, we observed that arterial enriched hiPSC-ECs formed a more extensive capillary network in this model, by comparison to a heterogeneous population of hiPSC-ECs. This study demonstrates that FACS purification of CD31+ hiPSC-ECs produces a diverse population of ECs. Refining the differentiation methods can enrich for subtype-specific hiPSC-ECs with functional benefits of enhancing neovascularization.
authors: Division of Cardiovascular Medicine, Stanford University School of Medicine 300 Pasteur Drive, Stanford, CA 94305, USA.
authors: Abdul Jalil
authors: AJ
authors: Rufaihah
authors: Ngan F
authors: NF
authors: Huang
authors: Jeanna
authors: J
authors: Kim
authors: Joerg
authors: J
authors: Herold
authors: Katharina S
authors: KS
authors: Volz
authors: Tea Soon
authors: TS
authors: Park
authors: Jerry C
authors: JC
authors: Lee
authors: Elias T
authors: ET
authors: Zambidis
authors: Renee
authors: R
authors: Reijo-Pera
authors: John P
authors: JP
authors: Cooke
keywords: Heterogeneity; angiogenesis; differentiation; endothelial cells; induced pluripotent stem cells
references: Vascul Pharmacol. 2012 May-Jun;56(5-6):288-96
references: 22387745
references: Immunol Today. 1994 Oct;15(10):490-5
references: 7945775
references: Circulation. 2010 Aug 3;122(5):517-26
references: 20679581
references: Nature. 2005 Dec 15;438(7070):937-45
references: 16355211
references: Proc Natl Acad Sci U S A. 1997 Jun 10;94(12):6273-8
references: 9177207
references: Nature. 2005 May 5;435(7038):98-104
references: 15875024
references: Histochem Cell Biol. 2008 Dec;130(6):1063-78
references: 18946678
references: Arterioscler Thromb Vasc Biol. 2010 May;30(5):984-91
references: 20167654
references: Development. 2001 Oct;128(19):3675-83
references: 11585794
references: Semin Thromb Hemost. 2010 Apr;36(3):227-35
references: 20490975
references: Circulation. 2008 Jul 29;118(5):498-506
references: 18625891
references: Blood. 2005 Jun 15;105(12):4642-8
references: 15746084
references: Circ Res. 2007 Feb 2;100(2):158-73
references: 17272818
references: Arterioscler Thromb Vasc Biol. 2007 Oct;27(10):2127-34
references: 17872458
references: Nat Rev Mol Cell Biol. 2006 May;7(5):359-71
references: 16633338
references: Curr Top Microbiol Immunol. 1993;184:37-45
references: 8313722
references: Arterioscler Thromb Vasc Biol. 2006 May;26(5):1073-8
references: 16543496
references: Dev Biol. 2010 May 1;341(1):196-204
references: 20193674
references: Nature. 1996 Apr 4;380(6573):439-42
references: 8602242
references: Stem Cells. 2009 Aug;27(8):1750-9
references: 19544443
references: Blood. 2006 Feb 1;107(3):1214-6
references: 16195336
references: Cell Adh Migr. 2007 Jul-Sep;1(3):133-6
references: 19262131
references: Genome Biol. 2008;9(12):243
references: 19138383
references: Arterioscler Thromb Vasc Biol. 2010 Jan;30(1):80-9
references: 19875721
references: Circ Res. 2009 Mar 13;104(5):576-88
references: 19286613
references: Nature. 1996 Apr 4;380(6573):435-9
references: 8602241
references: Circ Res. 2007 Feb 2;100(2):174-90
references: 17272819
references: Nature. 2000 Nov 2;408(6808):92-6
references: 11081514
references: J Cell Biol. 2010 Apr 19;189(2):325-38
references: 20404113
references: Circulation. 2007 Nov 20;116(21):2409-19
references: 17984381
references: Arterioscler Thromb Vasc Biol. 2011 Nov;31(11):e72-9
references: 21836062
references: Histol Histopathol. 2010 May;25(5):637-46
references: 20238301
references: Arterioscler Thromb Vasc Biol. 2007 Mar;27(3):487-93
references: 17185616
references: PLoS One. 2009 Dec 31;4(12):e8443
references: 20046878
references: Arterioscler Thromb Vasc Biol. 2006 Sep;26(9):1977-84
references: 16809546
references: Arterioscler Thromb Vasc Biol. 2006 Sep;26(9):2070-6
references: 16690875
references: Science. 1997 Dec 12;278(5345):1947-50
references: 9395397
references: Nat Rev Mol Cell Biol. 2007 Jun;8(6):464-78
references: 17522591
references: Dev Cell. 2002 Jul;3(1):127-36
references: 12110173