Hepatocyte Alpha-1 Antitrypsin Deficiency (A1ATD) - Definigen

A1AT disease modelled hepatocytes

DefiniGEN provide disease modelled alpha-1 antitrypsin deficiency hepatocytes which are highly functional patient-derived human hepatocytes.
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A1AT Hexagon
Product Overview

DefiniGEN provide disease modelled alpha-1 antitrypsin deficiency hepatocytes which are highly functional patient-derived human hepatocytes. The cells are generated using the dermal fibroblasts from an alpha-1 antitrypsin (A1AT) disorder patient carrying the ‘Z’ (E342K) point mutation in the A1AT gene SERPINA1. This mutation leads to the formation of mutant A1AT polymers that ultimately cause liver and lung damage. DefiniGEN’s A1AT patient derived hepatocytes represent an optimized disease model for drug discovery applications and are an effective tool for elucidating the underlying mechanisms of the disease.

  • Disease circuit verified ZZ E342K mutation in the SERPINA1 gene
  • Normal human genetics wild-type donor genetics and karyotype verified
  • Standardized cell product >98% human hepatocyte cells producing reproducible and relevant data
  • Optimized work flow we can deliver industrial quantities of cryopreserved cell products to fit client specification
Technical Data
Def-HEP A1ATD cell morphology

The Def-HEP A1ATD cells are patient representative human hepatocytes. When thawed and plated Def-HEP A1ATD cells form a monolayer with typical hepatocyte cobblestone morphology.

DefiniGEN A1AT cell morphology
Figure 1. Clear cobblestone morphology of Def-HEP A1ATD cells post-thaw.
Hepatocyte maturation markers

Def-HEP A1ATD cells display the functional characteristics of primary human hepatocytes (PHH) including albumin and A1AT production (Figure 2). Accordingly, they should be handled in a similar fashion to primary human hepatocytes.

Functional analysis DefiniGEN A1AT disease modelled hepatocytes
Figure 2. Functional analysis of hepatocyte-specific markers in Def-HEP A1ATD cells. Immunostaining of Def-HEP A1ATD cells overviewing the expression of general hepatocyte maturation markers.

Detection of disease markers via ELISA

ELISA for polymer and secreted A1AT using antibodies specific for A1AT polymers (2C1mAb, top) or all conformers of A1AT (bottom).

Detection of disease markers via ELISA DefiniGEN A1AT
Figure 3. Detection of disease specific markers in Def-HEP A1ATD cells. Quantification of intracellular A1AT polymers and intercellular total A1AT secretion in Def-HEP A1ATD cells. Primary human hepatocytes (PHH) and hepatocellular liver carcinoma cells (Hep G2) are used as internal controls.

Immunocytochemistry analysis of A1AT mutant polymer

Previous studies have shown that the Z allele (Glu342Lys) results in the formation of ordered polymers of a1-antitrypsin that are retained within the ER. This pathway of a1-antitrypsin polymerization is central to the clinical phenotype. We therefore used the 2C1 polymer specific monoclonal antibody to detect polymers within Def-HEP A1ATD hepatocytes. Polymers were detected by immunostaining (Figure 4).

DefiniGEN Immunocytochemistry A1AT mutant Polymer
Figure 4. Accumulation of a1-antitrypsin polymers in Def-HEP A1ATD cells. Immunostaining analyses for expression of misfolded polymeric a1-antitrypsin using the polymer-specific 2C1 antibody (green) or an antibody that detects all forms of a1-antitrpysin (red) in Def-HEP A1ATD disease modelled cells and control Def-HEP WT. Merged images are shown at right.