Hello everyone.
Now, I am isolating colonic fibroblasts from C57BL/6 mice, I have tried digestion with 0.05g/ml collagenase type 1, but very few fibroblasts are isolated.
Can any research colleagues give me some advice?
Try either
0.5mg collagenase A (Signa) /ml and 1.5 mg pancreatin/ml and incubated for 90 minutes at 37C in HBSS. While gentle shaking
Or
Make very small pieces of the tissue (2 mm approx), leave in a dish with culture medium with FBS and Pen/Step (do not dip tissue in the medium, change medium carefully from the side (remove old medium and add fresh medium) every 3 days. The fibroblasts will proliferate in 10 to 15 days, you should get monolayers in less than 15 days. It is slow but non-enzymatic method, I have done in many tissues, it worked
