Comparative Profiling of ASC Products Following Harvest with DisoNNex and TrypLE
Cytotherapy, 2026
Johansen E., Bangsgaard S., Lethager L., Højgaard L., Hu T., Bugge K., Wilkens C., Nørgaard M.
| Disease area | Application area | Sample type | Products |
|---|---|---|---|
Wider Proteomics Studies | Technical Evaluation | Cell Lysate |  Olink Target 96 |
Abstract
Background
Cell therapies based on adipose tissue-derived mesenchymal stromal cells (ASC) are promising candidates for treatment of numerous diseases. Relevant to the potential of these therapies is the development of consistent and efficient ASC manufacturing processes, with enzymatic cell dissociation being a critical step. DisoNNex, a novel cell dissociation enzyme based mainly on chymotrypsin activity, offers a Good Manufacturing Practice (GMP)-compliant alternative to existing dissociation methods; its impact on ASC characteristics remains yet to be explored.
Methods
DisoNNex performance was compared to the current dissociation industry standard, TrypLE Select 1x (TrypLE). The DisoNNex concentration matching dissociation kinetics of TrypLE was defined using timelapse microscopy and image analysis. ASCs from five donors were harvested using either TrypLE or DisoNNex across two passages. Assays assessed immunophenotype, apoptosis, secretion of vascular endothelial factor, Indoleamine 2,3-dioxygenase expression, and suppression of lymphocyte proliferation. Multi-omics profiling included bulk RNA sequencing, proteomics, metabolomics, and lipidomics.
Results
DisoNNex at 15 mg/mL matched TrypLE in dissociation kinetics and maintained ASC viability. No significant differences were observed in functional assays. Multi-omics analysis revealed only four differentially expressed genes out of 12509 protein coding genes, and no significant differences in proteins, metabolites, or lipids were found. Multi-Omics Factor Analysis plot showed no clustering by treatment.
Conclusion
DisoNNex is a relevant alternative to TrypLE for ASC harvest, resulting in equivalent yields and cell quality under the tested conditions. These findings support DisoNNex’ potential in GMP-compliant manufacturing of ASC based cell therapies, and provide starting conditions for its implementation in ASC processes. Nonetheless, full comparability studies within an established production process are pending.