Optimized human cell products for research
and drug discovery

Hypercholesterolemia Hepatocytes

DefiniGEN Familial Hypercholesterolemia Disease Modelled Human Hepatocytes

Familial hypercholesterolemia (FH) is an autosomal dominant disorder of lipoprotein metabolism caused mainly by mutations in the low-density lipoprotein receptor (LDLR) gene. Disease modelled Familial Hypercholesterolemia hepatocytes (Def-HEP FH) are highly functional human hepatocytes derived using human induced pluripotent stem cell (iPSC) technology.

For the production of Def-HEP FH cells, fibroblasts are first reprogrammed into iPSC using the Nobel Prize winning technology developed by Yamanaka and colleagues. HORIZON CRISPR gene-editing is then used to introduce a precise genetic mutation into the LDLR gene of an iPS cell line. Def-HEP FH hepatocytes represent an optimized disease model for drug discovery applications and a principal tool for elucidating the underlying mechanisms of the disease.

Product Specification

Product ID:
Def-HEP FH
Format:
Cryopreserved
Cell Number:
Typically 3-6 million cells per vial
Disease:
Familial Hypercholesterolemia
Viability:
>70%
Applications:
Disease Modelling and Drug Discovery

Genetic Validation and Cell Viability

Def-HEP FH cells have been validated and verified for the E101K genetic mutation in the LDLR gene. Typical viabilities of the thawed hepatocyte are >70% upon receipt.

Disease circuit verification

Sequence Confirmation of LDLR E101K mutation

Figure 1. Sequence confirmation of LDLR E101K mutation in Def-HEP FH cells.

Functional Test

The in vivo functional implications of LDL receptor deficiency are conserved in our model, as shown by immunostaining and FACS analysis demonstrating that Def-HEP FH iPS cell–derived hepatocytes have an impaired ability to incorporate LDL. Receptor-specific binding of DiI-LDL is followed by internalization of the bound complex and lysosomal hydrolysis of the ligand. Increase in the fluorescence intensity per cell is hence used as a measure of DiI-LDL uptake and, implicity, as an indication of LDL-R presence. These results demonstrate that CRISPR generated disease-specific human iPS cells can successfully be used to model FH.

LDL receptor deficiency

                 Wild Type                                    FH Disease Model

Figure 2. These microscopy images demonstrate that uptake of LDL was impaired in the LDLR mutant line in comparison to WT control. 

LDL Uptake

 

LDL Uptake

Figure 3. Quantitative LDL receptor assays based on fluorescent DiI-LDL internalization have demonstrated that Def-HEP FH iPS cell–derived hepatocytes have a significantly impaired ability to incorporate LDL relative to the isogenic control Def-HEP WT over a range of LDL substrate ranges.

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