Optimized human cell products for research
and drug discovery

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 Specification

Product ID:
Def-CHOLANGIOCYTE WT
Format:
Cryopreserved
Application:
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

Organoid System

Cystic structure Branching (arrow) tubular structure

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.

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 Analysis

Immunofluorescence analyses, revealing much lower CFTR expression in CF-CLC than in WT-CLCs expressing CFTR. Scale bars, 100um.

Immunofluorescence Analysis

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.

CFTR Function VX809

Figure 2. Incubation of CF-CLCs with VX809 for 48hr increased CFTR function analyzed by MQAE to a level similar to what of WT-CLCs. This effect was negated by CFTR inhibitor 172, confirming that the phenotypic rescue of CF-CLCs by VX809 depended on improved CFTR function.