Derivation of Brain Capillary-like Endothelial Cells from Human Pluripotent Stem Cell-Derived Endothelial Progenitor Cells
| Autor: | Emmanuel Sevin, Marie-Pierre Dehouck, Catarina Praça, Lino Ferreira, Roméo Cecchelli, S.C. Rosa |
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| Přispěvatelé: | University of Coimbra [Portugal] (UC), Universidade de Coimbra [Coimbra], Laboratoire de la Barrière Hémato-Encéphalique (LBHE), Université d'Artois (UA) |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Pluripotent Stem Cells induced pluripotent stem cells [SDV]Life Sciences [q-bio] Biology Blood–brain barrier Biochemistry Immunophenotyping Extracellular matrix 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Report Genetics medicine Humans Progenitor cell blood-brain barrier brain capillary-like endothelial cells extracellular matrices in vitro model soluble factors Induced pluripotent stem cell Cells Cultured Endothelial Progenitor Cells Lucifer yellow Cell Differentiation Cell Biology Cell biology Extracellular Matrix Fibronectin Endothelial stem cell 030104 developmental biology medicine.anatomical_structure chemistry Blood-Brain Barrier biology.protein Endothelium Vascular Signal transduction 030217 neurology & neurosurgery Biomarkers Developmental Biology |
| Zdroj: | Stem Cell Reports Stem Cell Reports, Elsevier, 2019, 13 (4), pp.599-611. ⟨10.1016/j.stemcr.2019.08.002⟩ Repositório Científico de Acesso Aberto de Portugal Repositório Científico de Acesso Aberto de Portugal (RCAAP) instacron:RCAAP |
| ISSN: | 2213-6711 |
| Popis: | Summary The derivation of human brain capillary endothelial cells is of utmost importance for drug discovery programs focusing on diseases of the central nervous system. Here, we describe a two-step differentiation protocol to derive brain capillary-like endothelial cells from human pluripotent stem cells. The cells were initially differentiated into endothelial progenitor cells followed by specification into a brain capillary-like endothelial cell phenotype using a protocol that combined the induction, in a time-dependent manner, of VEGF, Wnt3a, and retinoic acid signaling pathways and the use of fibronectin as the extracellular matrix. The brain capillary-like endothelial cells displayed a permeability to lucifer yellow of 1 × 10−3 cm/min, a transendothelial electrical resistance value of 60 Ω cm2 and were able to generate a continuous monolayer of cells expressing ZO-1 and CLAUDIN-5 but moderate expression of P-glycoprotein. Further maturation of these cells required coculture with pericytes. The study presented here opens a new approach for the study of soluble and non-soluble factors in the specification of endothelial progenitor cells into brain capillary-like endothelial cells. Highlights • Derivation of BCLECs from iPSCs in chemically defined medium in two steps • Specification of EPCs into BCLECs requires the activation of VEGF, Wnt3a, and RA • Fibronectin seems a sufficient substrate for the specification of EPCs into BCLECs In this article, Ferreira and colleagues describe a protocol for the derivation of brain capillary-like endothelial cells from iPSCs under fully chemically defined medium and extracellular matrix in two steps. The protocol requires the modulation of three signaling pathways (VEGF, Wnt3a, and retinoic acid) and the use of fibronectin as a single extracellular matrix component. |
| Databáze: | OpenAIRE |
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