Hi, we are trying to isolate fibroblasts from human colon biopsies (from UC patients) using enzymatic digestion with collagenase type IV for 40min, followed by mechanical digestion using gentlemacs. Cells are passed through filter to get rid off tissue debris, spun down and then cultured in DMEM+16% FCS. So far we havent been very successful so I am wondering if anyone has a protocol that is working. Thanks in advance.