Directions for Use

Download the Directions for Use PDF or continue reading below:

1. Use these recommendations as guidelines to determine the optimal gel dissociation procedure for your culture system.
2. Reconstitute the lyophilized collagenase powder (50 mg) in phosphate buffered saline or other pH neutral medium. A typical concentration of 1 mg/ml may be used.
3. Aspirate the culture medium from the collagen gel.
4. Wash the collagen gel 2X with pre-warmed PBS or other pH neutral medium to remove culture medium which could inhibit the degradation of collagen.
5. Add appropriate amount of collagenase solution (one gel volume should be sufficient). For example: For 24 well plates, containing 0.5 ml of gel, add 0.5 ml of collagenase solution.
6. Transfer to a 37ºC incubator for 30-60 minutes. To facilitate the faster dissociation of the collagen gel, the gel and collagenase solution can be pipetted up and down using a large bore pipette tip.
7. Incubate additional 30-60 minutes at 37°C, if necessary, to complete the digestion of the gel. If required, pipette the gel/cell mixture every 15 minutes. Note: Thicker gels or gels containing higher concentration of cells may require more time.
8. Once the gel is fully digested, add an equal volume of complete culture medium to the gel/cell mixture and then collect the cells.
9. Rinse the culture vessel collecting any residual cells.
10. Centrifuge the cell suspension at 250 x g for 5 minutes at room temperature.
11. Carefully aspirate the supernatant, and resuspend the cell pellet in 0.5 -1 ml of fresh medium.
12. Determine cell count and viability using a hemocytometer and Trypan Blue.

Product Q & A

Product References

Collagenase References:

Röck, Katharina, et al. "Collagen fragments inhibit hyaluronan synthesis in skin fibroblasts in response to ultraviolet B (UVB) new insights into mechanisms of matrix remodeling." Journal of Biological Chemistry 286.20 (2011): 18268-18276.

Shieh, Hester F., et al. "Comparisons of human amniotic mesenchymal stem cell viability in FDA-approved collagen-based scaffolds: Implications for engineered diaphragmatic replacement." Journal of pediatric surgery 52.6 (2017): 1010-1013.

Kumar, Balasubramanian K., et al. "The effects of over expressing aquaporins on the cryopreservation of hepatocytes." Cryobiology 71.2 (2015): 273-278.

Yang, Xi, and William F. Salminen. "Kava extract, an herbal alternative for anxiety relief, potentiates acetaminophen-induced cytotoxicity in rat hepatic cells." Phytomedicine 18.7 (2011): 592-600.

Ingeson-Carlsson, Camilla, Angela Martinez-Monleon, and Mikael Nilsson. "Differential effects of MAPK pathway inhibitors on migration and invasiveness of BRAFV600E mutant thyroid cancer cells in 2D and 3D culture." Experimental cell research 338.2 (2015): 127-135.

Voedisch, Sabrina, et al. "Neuropeptides control the dynamic behavior of airway mucosal dendritic cells." PloS one 7.9 (2012): e45951.

Safety and Documentation

Safety Data Sheet

Certificate of Origin

Product Disclaimer

This product is for R&D use only and is not intended for human or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.