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
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.
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.
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).
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).