Human Cholangiocytes Wild Type – Coming Soon
DefiniGEN Cholangiocyte Organoids
Def-Cholangiocyte cells are a specialized cell product similar in performance and function to primary cholangiocyte cells. Accordingly the cell products show the functional characteristics of cholangiocytes, including bile acids transfer, alkaline phosphatase activity, glutamyl-transpeptidase activity and physiological responses to secretin, somatostatin and vascular endothelial growth factor. The cell products grow as cystic organoids and branching tubular structures bearing primary cilia mimicking physiological biliary development, generating cell populations that closely resemble primary human cholangiocytes at the transcriptional and functional level. The products can be used as an optimized in vitro system to model key features of Alagille syndrome, polycystic liver disease and cystic fibrosis (CF)-associated cholangiopathy. Furthermore, wild-type and CF disease modelled Def-Cholangiocyte products generated from healthy individuals and patients with polycystic liver disease can be used to reproduce the effects of the drugs verapamil and octreotide, and show that the experimental small molecule CF drugs can rescue the disease phenotype of CF cholangiopathy in vitro.
- Product ID:
- Def-CHOLANGIOCYTE WT
- Research and Drug Discovery
Display key functional characteristics including bile acids transfer, alkaline phosphatase activity, response to hormonal stimuli and CFTR activity
Organoid system which provides a unique in vitro system to model human liver cholangiocyte cells
Effectively undertake CF modelling it has been demonstrated that CFTR-Df508 mis-folded receptor can be rescued to a functional state in cholangiocyte organoids
Figure 1. Immunofluorescence images of Cholangiocyte organoids demonstrating the formation of cystic (a) and branching (arrows) tubular structures (b). Scale bars, 100um.
Expression of Key Biliary Markers
QPCR analysis of Cholangiocyte organoids demonstrated the expression of key biliary markers.
Figure 2. QPCR analyses of CLC organoids generated from CF-hiPSCS, demonstrating the expression of biliary markers. Astericks denote statistical significance in differences between HBs, CPs and CLCs (one-way ANOVA with Tukey correction for multiple comparisons). Values are relative to the housekeeping gene PBGD. Center line, median; box, interquartile range (IQR)l whiskers, range (minimum to maximum).
Immunofluorescence analyses, revealing much lower CFTR expression in CF-CLC than in WT-CLCs expressing CFTR. Scale bars, 100um.
Figure 1. Immunofluorescence analyses detected minimal CFTR protein expression.
CFTR Function – VX809
MQAE fluorescence intensity, normalized to the lowest intensity value. MQAE fluorescence is quenched in the presence of chloride but not affected by nitrate. Changes in intracellular or intraluminal chloride in response to extracellular chloride changes depend on the presence of CFTR functionality. MQAE fluorescence increases in response to a nitrate challenge depleting extracellular chloride and decreases in response to chloride in WT – and CF-CLCs treated with VX809, but fails to respond to both challenges in CF-CLCs and CF-CLCs treated with VX809 plus CFTR inhibitor 172. Error bars represent s.d.
CLC organoids appropriately modified intracellular cholride in response to media with varying chloride concentrations, whereas no change was observed in CF-CLCs, confirming the absence of functional CFTR in these cells.