Rat ventral prostate cell lysis at 2 degrees C.
In order to physically separate epithelial from nonepithelial cells, well-minced rat ventral prostate at 2 degrees C was passed through a tissue sieve, and the disrupted tissue suspended and washed several times before centrifugation on a Ficoll gradient. While limited separation of single prostate epithelial and nonepithelial cells and small aggregates could be achieved, the yield of intact undamaged cells was low, and many nuclei contaminated the 2 major cell fractions obtained from the gradient. During subsequent experiments, it became apparent that most single cells released from prostates minced at 2 degrees C rapidly lysed, yielding cytoplasmic debris and cell nuclei. Yet the morphology of red and white blood cells, examined by light microscopy, was unaffected, suggesting a soluble factor was not responsible. These results indicated that mechanical dissociation of rat ventral prostate at 2 degrees C with release of single cells was accompanied by a powerful prostate cell-associated lytic 'event', affecting both epithelial and connective tissue cells, without destruction of cell nuclei or accompanying red and white blood cells. Some properties of this process are described in this report.
D000818:Animals; D002453:Cell Cycle; D002458:Cell Fractionation; D002467:Cell Nucleus; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D002498:Centrifugation; D002860:Chromium Radioisotopes; D003080:Cold Temperature; D008297:Male; D003012:Microbial Collagenase; D011467:Prostate / Q000166:cytology*; D051381:Rats; D011919:Rats, Inbred Strains; D014343:Trypan Blue
D002860:Chromium Radioisotopes; D003012:Microbial Collagenase; D014343:Trypan Blue