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Monoclonal antibodies against cell surface antigens expressed during porcine adipocyte differentiation
1U.S. Department of Agriculture, Animal Physiology Research Unit, Richard B. Russell Agricultural Research Center, Athens, Georgia 30613.
Two monoclonal antibodies (AD-1 and AD-2) were prepared by fusion of mouse myeloma cells and lymph node cells of mice immunized with porcine adipocyte plasma membranes. Immunoprecipitation of iodinated adipocyte plasma membrane proteins followed by SDS-PAGE and autoradiography yielded protein antigens for each antibody. The AD-1 and AD-2 antigens were detected on mature adipocytes and a proportion of non-lipid-containing cells in stromal-vascular cultures. Adipocytes and associated capillary networks in subcutaneous adipose tissues as well as capillaries between the underlying muscle fiber bundles bound each antibody, whereas the AD-2 monoclonal antibody also reacted with vessels but not capillaries in liver tissues. In stromal-vascular cell cultures prepared from newborn pig subcutaneous tissue, the AD-1 and AD-2 antibodies exhibited reactivity towards 45 percent and 10 percent respectively, of cells 24 hours after seeding. On the other hand, only 4 percent and 1 percent of the cells in cultures prepared from 60 day fetal subcutaneous tissues expressed detectable amounts of the AD-1 and AD-2 antigens, respectively. In conclusion, cells along the adipogenic lineage possess cell surface antigens which may not be unique to adipogenic cells, but do exhibit differential expression among cell populations within adipose tissues. A temporal relationship between adipogenesis and angiogenesis was also demonstrated.
Two monoclonal antibodies (AD-1 and AD-2) were prepared by fusion of mouse myeloma cells and lymph node cells of mice immunized with porcine adipocyte plasma membranes. Immunoprecipitation of iodinated adipocyte plasma membrane proteins followed by SDS-PAGE and autoradiography yielded protein antigens for each antibody. The AD-1 and AD-2 antigens were detected on mature adipocytes and a proportion of non-lipid-containing cells in stromal-vascular cultures. Adipocytes and associated capillary networks in subcutaneous adipose tissues as well as capillaries between the underlying muscle fiber bundles bound each antibody, whereas the AD-2 monoclonal antibody also reacted with vessels but not capillaries in liver tissues. In stromal-vascular cell cultures prepared from newborn pig subcutaneous tissue, the AD-1 and AD-2 antibodies exhibited reactivity towards 45 percent and 10 percent respectively, of cells 24 hours after seeding. On the other hand, only 4 percent and 1 percent of the cells in cultures prepared from 60 day fetal subcutaneous tissues expressed detectable amounts of the AD-1 and AD-2 antigens, respectively. In conclusion, cells along the adipogenic lineage possess cell surface antigens which may not be unique to adipogenic cells, but do exhibit differential expression among cell populations within adipose tissues. A temporal relationship between adipogenesis and angiogenesis was also demonstrated.