Differentiation of Human Embryonic Stem Cells to Endothelial Progenitor Cells on Laminins in Defined and Xeno-free Systems.
A major hurdle for in vitro culturing of primary endothelial cells (ECs) is that they readily dedifferentiate, hampering their use for therapeutic applications. Human embryonic stem cells (hESCs) may provide an unlimited cell source; however, most current protocols deriving endothelial progenitor cells (EPCs) from hESCs use direct differentiation approaches albeit on undefined matrices, yet final yields are insufficient. We developed a method to culture monolayer hESCs on stem cell niche laminin (LN) LN511 or LN521 matrix. Here, we report a chemically defined, xeno-free protocol for differentiation of hESCs to EPCs using LN521 as the main culture substrate. We were able to generate ∼95% functional EPCs defined as VEGFR2+CD34+CD31+VE-Cadherin+. RNA-sequencing analyses of hESCs, EPCs, and primary human umbilical vein endothelial cells showed differentiation-related EC expression signatures, regarding basement membrane composition, cell-matrix interactions, and changes in endothelial lineage markers. Our results may facilitate production of stable ECs for the treatment of vascular diseases and in vitro cell modeling.
Cardiovascular and Metabolic Disorders Program, Duke-NUS Medical School, Singapore 169857, Singapore.
Mien T X
MTX
Nguyen
Cardiovascular and Metabolic Disorders Program, Duke-NUS Medical School, Singapore 169857, Singapore.
Elena
E
Okina
Cardiovascular and Metabolic Disorders Program, Duke-NUS Medical School, Singapore 169857, Singapore.
Xiaoran
X
Chai
Division of Obstetrics and Gynaecology, KK Women's & Children's Hospital, Singapore 229899, Singapore.
Kok Hian
KH
Tan
Department of Clinical Sciences, Intervention and Technology, Karolinska Institute, Karolinska University Hospital Huddinge, Stockholm 14186, Sweden.
Outi
O
Hovatta
Cardiovascular and Metabolic Disorders Program, Duke-NUS Medical School, Singapore 169857, Singapore.
Sujoy
S
Ghosh
Cardiovascular and Metabolic Disorders Program, Duke-NUS Medical School, Singapore 169857, Singapore; Division of Matrix Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm 17177, Sweden. Electronic address: karl.tryggvason@duke-nus.edu.sg.
Karl
K
Tryggvason
D015415:Biomarkers; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D016000:Cluster Analysis; D066026:Endothelial Progenitor Cells / Q000166:cytology* / Q000378:metabolism*; D004727:Endothelium / Q000196:embryology / Q000378:metabolism; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D000066449:Human Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D007797:Laminin / Q000378:metabolism*; D008648:Mesoderm / Q000196:embryology / Q000378:metabolism; D015398:Signal Transduction; D059467:Transcriptome
D015415:Biomarkers; D007797:Laminin
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