Flow cytometric analysis of porcine preadipocyte replication.
In the present study, a technique is described for measuring proliferation of primary porcine preadipocytes. Cultures of stromal-vascular (S-V) cells from dorsal subcutaneous adipose tissue were maintained under several growth conditions. Cells were isolated from cultures at various times and DNA was labeled with propidium iodide (PI) and analyzed by flow cytometry. Treatment of cultures with colchicine allowed identification of S-V cells in mitosis. The relative proportion of replicating cells was dependent on several factors, including serum concentration and cell density at the time of harvest. Using the AD-1 anti-preadipocyte/adipocyte monoclonal antibody, the preadipocyte subpopulation within the S-V cells was identified. Furthermore, by labeling cells with PI concomitant with AD-1, the proportion of replicating preadipocytes was quantified. It is now possible to identify and quantify replicating preadipocytes rapidly under various experimental conditions.
Wright
J T
JT
Animal Physiology Unit, R. B. Russell Research Center, ARS, USDA, Athens, GA 30613.
Hausman
G J
GJ
D017667:Adipocytes / Q000166:cytology* / Q000276:immunology; D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal / Q000032:analysis; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D003470:Culture Media; D004247:DNA / Q000032:analysis; D005434:Flow Cytometry / Q000662:veterinary; D016650:Fluorescein-5-isothiocyanate; D011419:Propidium; D013552:Swine / Q000254:growth & development*
D000911:Antibodies, Monoclonal; D003470:Culture Media; D011419:Propidium; D004247:DNA; D016650:Fluorescein-5-isothiocyanate