Isolated adipocytes: an assessment of cell surface changes during their preparation.
A method is described, based on the detection of adipocyte-specific cell surface antigens, which allows assessment of the relative surface damage incurred by the cells when they are prepared under a variety of conditions. Using the method it is possible to develop, for any set of reagents, a set of cell isolation conditions (collagenase concentration, time of incubation) which will produce minimally damaged cells which exhibit high levels of specific cell surface immunoreactivity. Under certain conditions a recovery from limited surface damage can be achieved, although, when cells are prepared under more extreme conditions irreversible surface damage occurs. The surface morphology of the cells as revealed by scanning electron microscopy, is also clearly affected by the conditions of cell isolation. The method has been used to define the conditions necessary for the isolation of cells to be used in the study of subtle biochemical responses.
D000273:Adipose Tissue / Q000032:analysis / Q000166:cytology*; D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D002458:Cell Fractionation / Q000379:methods; D002462:Cell Membrane / Q000032:analysis / Q000648:ultrastructure; D002469:Cell Separation / Q000379:methods; D002499:Centrifugation, Density Gradient; D004822:Epididymis; D066298:In Vitro Techniques; D008297:Male; D003012:Microbial Collagenase; D051381:Rats; D011919:Rats, Inbred Strains
D000954:Antigens, Surface; D003012:Microbial Collagenase