human female ips imr90 4 wisci004 b  (WiCell Research Institute Inc)

Journal: Frontiers in Cell and Developmental Biology

Article Title: Using a brain-like endothelial cell differentiation to characterize the CS79iBRCA-n2 BRCA1 mutated patient derived stem cell line

doi: 10.3389/fcell.2025.1516669

Figure Lengend Snippet: CS79iBRCA-n2-derived BECs have endothelial properties. (A) Graphic outlining the process of harvesting the hiPSCs and their differentiation into BECs. (B) Timeline outlining the differentiation process. (C) qPCR showing expression of pluripotency marker, POUF51 , in CS79-hiPSCs compared to CS79-derived BECs ( N = 6). (D) TEER measurements from CS79-derived BECs over a period of 8 days post purification from four independent differentiations conducted in triplicate per differentiation ( n = 12). TEER measurements of three CS79-hiPSCs passages over a period of 5 days post passaging. (E) TEER measurements from IMR90-4 derived BECs over a period of 8 days post purification for three independent differentiations ( n = 9). (F) Representative immunofluorescence images of CS79-hiPSCs and BECs for endothelial markers VE-Cadherin and PECAM-1 (green) with nuclei stained with DAPI (blue). (G) Representative immunofluorescence images of IMR90-4 derived BECs for endothelial markers VE-Cadherin and PECAM-1 (green) with nuclei stained with DAPI (blue). (H) Western blot analysis of CS79-hiPSCs and CS79-derived BECs probing for endothelial marker VE-Cadherin with densitometry quantification. PonS stain shown to show relative protein concentrations loaded for each lane ( n = 3). (I) Western blot analyses of IMR90-4 hiPSCs and IMR90-4 derived BECs probing for endothelial marker VE-Cadherin with densitometry quantification. PonS stain shown to show relative protein concentrations loaded for each lane ( n = 3). Scale bar = 50 μm. Statistical significance calculated by Student’s t -test, ** p < 0.01, **** p < 0.0001. Error bars represent SD.

Article Snippet: The IMR90-4 cell line was purchased from WiCell (WISCi004-B) where it was reprogrammed using viral transfection methods from a female parent cell line, IMR90.

Techniques: Derivative Assay, Expressing, Marker, Purification, Passaging, Immunofluorescence, Staining, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: Using a brain-like endothelial cell differentiation to characterize the CS79iBRCA-n2 BRCA1 mutated patient derived stem cell line

doi: 10.3389/fcell.2025.1516669

Figure Lengend Snippet: Tight junction expression of CS79iBRCA-n2-derived BECs. (A) Representative immunofluorescence images of CS79-hiPSCs and CS79-derived BECs for the tight junction proteins, ZO-1, Occludin, and Claudin-5 (green) and nuclei stained with DAPI (blue). Scale bar = 50 μm. (B) Representative immunofluorescence images of IMR90-4 derived BECs for tight junction proteins Claudin-5, Occludin, and ZO-1. Scale bar = 50 μm. (C) Western blot analyses of CS79-hiPSCs and CS79-derived BECs probing for tight junction proteins, ZO-1, Occludin, and Claudin-5. PonS stain shown to show relative protein concentrations loaded for each lane ( n = 3). Quantification of western blots for ZO-1, Occludin, and Claudin-5 ( n = 3). (D) Western blot analyses of IMR90-4 hiPSCs and IMR90-4 derived BECs probing for ZO-1 and Occludin. PonS stain shown to show relative protein concentrations loaded for each lane ( n = 3). Quantification of western blots for ZO-1, Occludin, and Claudin-5 ( n = 3). Statistical significance calculated by Student’s t -test, * p < 0.05, ** p < 0.01. Error bars represent SD.

Article Snippet: The IMR90-4 cell line was purchased from WiCell (WISCi004-B) where it was reprogrammed using viral transfection methods from a female parent cell line, IMR90.

Techniques: Expressing, Derivative Assay, Immunofluorescence, Staining, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: Using a brain-like endothelial cell differentiation to characterize the CS79iBRCA-n2 BRCA1 mutated patient derived stem cell line

doi: 10.3389/fcell.2025.1516669

Figure Lengend Snippet: Characterization of the expression and localization of transporter proteins in CS79iBRCA-n2-derived BECs. (A) Representative immunofluorescence images for CS79-hiPSCs and CS79-derived BECs for nutrient transporter GLUT1, efflux transporter BCRP, and efflux transporter P-gp (green) and nuclei stained with DAPI (blue). Scale bar = 50 μm. (B) Representative immunofluorescence images for IMR90-4 derived BECs expression of GLUT1, BCRP, and P-gp (green) and nuclei staining with DAPI (blue). Scale bar = 50 μm. Substrate accumulation assays determining the function of the efflux transporters (C) BCRP and (D) P-gp for three independent differentiations conducted in triplicate ( n = 9). (E) Uptake assay determining large molecule transport of 10 kD Dextran in CS79-hiPSCs and CS79-derived BECs ( n = 9). (F) Sodium fluorescein transport assay in CS79-hiPSCs and CS79-derived BECs ( n = 9). Student’s t -test was used to determine statistical significance within the same cell conditions, **** p < 0.0001. Error bars represent SD.

Article Snippet: The IMR90-4 cell line was purchased from WiCell (WISCi004-B) where it was reprogrammed using viral transfection methods from a female parent cell line, IMR90.

Techniques: Expressing, Derivative Assay, Immunofluorescence, Staining, Transport Assay