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. Author manuscript; available in PMC: 2020 Jul 1.
Published in final edited form as: Arterioscler Thromb Vasc Biol. 2019 May 9;39(7):1317–1329. doi: 10.1161/ATVBAHA.119.312265

Table 1.

Key advances in various techniques for deriving ECs from various cell sources.

 Co-culture Initial cell type Feeder cell type EC differentiation Key advance Refs
hESCs Mouse bone marrow stromal cells (S17 or M2–10B4)

  1. Sort CD34+ cells

  2. Culture in EC media

 First indication that ECs may be generated from hESC-derived hematoendothelial precursors.

  1. Kaufman PNAS 2001

  2. Hill Exp Hematol. 2010
 Embryoid body Initial cell type EC differentiation Key advance Refs
hESCs

  1. Sort CD31+ cells from dissociated EBs

 First study to show that ECs could be derived from spontaneously differentiating EBs. Levenberg PNAS 2002
hESCs

  1. Treat EBs with hematopoetic cytokines and BMP4

  2. Sort VE-Cad+/CD31+/VEGFR2+/CD45− cells

  3. Culture in EC media

 First study to use mesodermal cues to direct differentiation of EBs to hematoendothelial precursor fate.

  1. Chadwick Blood 2003

  2. Wang Immunity 2004
hESCs

  1. Treat EBs with BMP4/FGF2/Activin A to form primitive streak

  2. Treat with VEGF/DKK1 to specify cardiogenic mesoderm

  3. Treat with VEGF/DKK1/FGF2 to specify ECs

 Showed that ECs can be derived from multipotent cardiovascular progenitor cells by directing differentiation of EBs to cardiogenic mesoderm. Yang Nature 2008
hESCs

  1. Treat EBs with BMP4/Activin A/FGF2/VEGF to specify mesoderm and ECs

  2. Treat with TGFβ inhibitor to expand ECs

 Inhibiting TGFβ can improve EC yield and maintain EC identity. James Nat Biotechnol. 2010
 Monolayer Initial cell type EC differentiation Key advance Refs
hESCs/
hiPSCs

  1. Treat with CHIR

  2. Sort for CD34+ cells

  3. Culture in EC media

 Demonstrated that a small molecular GSK3 inhibitor (CHIR), rather than cytokines, can be used to generate cardiovascular progenitor cells. Lian Stem Cell Reports 2014
hESCs/
hiPSCs

  1. Treat with Activin A/BMP4/FGF2 to induce mesodermal formation

  2. Sort for NRP1+CD31+ cells

  3. Treat with FGF2/BMP4/VEGF to specify ECs

 Generated cord-blood endothelial colony-forming cells with high proliferative potential. Prasain Nat Biotechnol. 2014
hESCs/
hiPSCs

  1. Treat with GSK3 inhibitor & BMP-4 to induce mesoderm formation

  2. Treat with VEGF & PKA activator to specify ECs

 Faster and more efficient monolayer-based EC generation. Patsch Nat Cell Biol. 2015
hESCs/
hiPSCs

  1. Titrate Activin A/BMP4 and add CHIR to derive either cardiogenic and hemogenic mesoderm

  2. Treat with BMP4/FGF2/VEGF to specify ECs

  3. Treat with FGF2/VEGF/GSK3 inhibitor to mature ECs

 Generated developmentally-distinct ECs by patterning anterior and posterior-like mesoderm.

  1. Palpant Development 2015

  2. Palpant Nat Protoc. 2017
 Transdifferentiation Initial cell type Reprogramming EC differentiation Key advance Refs
Amniotic Cells ETV2/FLI1/ERG1 lentivirus + TGFβ inhibitor

  1. Culture in endothelial media + TGFβ inhibitor

 Forced expression of key EC transcription factors reprograms amniotic cells to ECs. Ginsberg Cell 2012
Fibroblasts Sox2/Oct4/KLF4/
cMyc/LIN28/shP53 episomal vector

  1. Treat with FGF2/VEGF/BMP4 to induce mesoderm

  2. Culture in EC media to specify ECs

 Brief non-integrating expression of pluripotency factors combined with mesodermal induction can be used to generate angioblasts and subsequently, ECs. Kurian Nat Methods 2013

hESC – human embryonic stem cell, hiPSC – human induced pluripotent stem cell, EC – endothelial cell, EB – embryoid body.