Differentiated cell population of endothelial cells derived from human pluripotent stem cells, composition, system, kit and uses thereof

Abstract

The present disclosure relates to a differentiated cell population of endothelial cells derived from human pluripotent stem cells. The present invention also relates to a composition, a system and a kit comprising those cells and uses thereof. The present disclosure also described the combination of arterial ECs derived from human pluripotent stem cells with a microfluidic system to create a vascular kit for high-throughput drug screening and/or toxicology analysis. This technology may find particular use for the identification of drugs that may have a fetal cytotoxic effect.

1 . A differentiated cell population of endothelial cells derived from human pluripotent stem cells wherein a portion of the said endothelial cells express ephrin B2. 
     
     
         2 . The cell population according to  claim 1  wherein at least 20% of the cells express ephrin B2. 
     
     
         3 . The cell population according to  claim 1  wherein 50% to 75% of the cells express ephrin B2. 
     
     
         4 . The cell population according to  claim 1  wherein the said endothelial cells further express the following marker Ac-LDL. 
     
     
         5 . The cell population according to  claim 1  wherein the said endothelial cells further express one of the following markers: vWF, CD31, CD34, vascular endothelial cadherin, Flk-1/KDR. 
     
     
         6 . The cell population according to  claim 1  wherein the said endothelial cells express one the following arterial endothelial cell genes: jagged 1, jagged 2, ephrin B1, Hey-2. 
     
     
         7 . The cell population according to  claim 1  wherein the endothelial cells express one of the following markers: receptor protein tyrosine phosphatase, T-cell acute lymphocyte leukemia, N-cadherin, angiopoietin 1, DNA-binding protein inhibitor ID-1. 
     
     
         8 . The cell population according to  claim 1  wherein the said endothelial cells have a low expression of venous genes such as EphB4, lefty-A and lefty-B. 
     
     
         9 . A differentiated cell population of endothelial cells derived from human pluripotent stem cells wherein a portion of the said endothelial cells express: receptor protein tyrosine phosphatase, T-cell acute lymphocyte leukemia, N-cadherin, angiopoietin 1, DNA-binding protein inhibitor ID-1 or least one of the following markers: vWF, CD31, CD34, vascular endothelial cadherin (VE-CAD), Flk-1/KDR for use in screening embryonic vascular toxicity or therapeutic compounds, preferentially a portion of at least 25%. 
     
     
         10 . The cell population according to  claim 1  for use in medicine. 
     
     
         11 . The cell population according to  claim 1  for use in screening embryonic vascular toxicity or therapeutic compounds. 
     
     
         12 . (canceled) 
     
     
         13 . A composition comprising the cell population described in  claim 1 . 
     
     
         14 . A kit for use in screening vascular toxicity or therapeutic compounds comprising the cell population described in  claim 1 . 
     
     
         15 . A fluidic system for use in screening therapeutic drugs or embryonic vascular toxicity, comprising a channel with a millimeter or micrometer dimension and a differentiated cell population of endothelial cells as described in  claim 1  wherein the cells are cultured under physiologic shear stress. 
     
     
         16 . The system according to  claim 1  wherein the cells cultured under physiologic shear stress are cells seeded and exposed to the media flow. 
     
     
         17 . The system according to  claim 15  wherein the said cells produce glycocalyx. 
     
     
         18 . The system according to  claim 15  wherein the channel comprises poly(dimethylsiloxane). 
     
     
         19 .- 26 . (canceled) 
     
     
         27 . A device for use in screening vascular toxicity or therapeutic compounds comprising:
 endothelial cells derived from human pluripotent stem cells according to  claim 1 ;   and a fluidic system.   
     
     
         28 .- 32 . (canceled) 
     
     
         33 . A method of promoting the viability of arterial endothelial cells comprising differentiation from human pluripotent stem cells with the following steps:
 a) culturing embryoid bodies in suspension for 4 days in differentiation medium comprising VEGF 6 s;   b) seeding the said embryoid bodies in a cell culture and sequentially exposing the cells to VEGF plus thymosin β4 for 3 days and VEGF plus thymosin β for 4 days and SB431542 for 11 days;   c) isolating CD31+ cells and culturing these cells in endothelial cell medium containing SB431542.   
     
     
         34 .- 41 . (canceled)