Hello everyone. A colleague of mine has asked me to post his question to the all knowing flow community: We've been trying to harvest mesothelial cells from the peritoneum of mice with very limited success. A couple of protocols we've used has involved: 1. Injecting warm trypsin(0.25%)/EDTA into the peritoneal cavity and harvesting the cells after ten minutes. We've only been able to get about 20,000 mesothelial cells per mouse (they seem to grow happily though). 2. Digesting chopped up peritoneal tissue with trypsin. There were very few cells harvested and most were dead. 3. Digesting chopped up peritoneal tissue with 0.1% collagenase/DNAse. Hardly any cells and they were mostly dead. Does anyone out there in the big wide flow world have any good suggestions for harvesting murine peritoneal mesothelial cells for culture. Any input would be greatly appreciated. Kylie Price Flow Cytometry Manager The Malaghan Institute of Medical Research The Central Services Building Victoria University Campus Entrance 7 Kelburn Parade Wellington 6242 New Zealand Ph: 64-4-4996914, ext 850 Fax: 64-4-4996915 E-mail: kprice@malaghan.org.nz Web: www.malaghan.org.nz
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