Table 1 Troubleshooting common issues encountered during tissue dissociation for intestinal crypt isolation via semi-automated system.
From: Automated tissue dissociation for the establishment of human intestinal organoids
Problem | Solution |
|---|---|
Biopsy adhesion to Via Extractor Sachet / pipette tips | Pre-coat plastic containers and equipment with 0.1% BSA prior to addition of biopsies |
Organoids fail to grow from the dissociation mixture | 1. Ensure crypts are fully released and tissue is not under-dissociated (but avoid over dissociation) |
2. Minimise exposure to EDTA solution | |
3. Maintain dissociation temperature at 4 °C | |
Organoids show impaired growth | Adjust dissociation parameters to balance cell yield and viability, ensuring crypts (not single cells) are isolated |
