Production of heparin-binding endothelial mitogens by bovine uterine fibroblastic and epithelial cells.
This study was conducted to determine whether early-passage cultures of bovine endometrial fibroblastic (Fb, n = 7 uteri) and epithelial (Ep, n = 3 uteri) cells produce endothelial mitogens in vitro and to begin characterization of these mitogens. Confluent cultures of Fb and Ep were incubated for 72 h in serum-free media, and the resulting conditioned media (CM) were evaluated for effects on proliferation of bovine aortic endothelial cells. CM from these Fb cultures (n = 8) and Ep cultures (n = 4) stimulated (147 +/- 10% (mean +/- SEM), p < 0.01, and 124 +/- 8%, p < 0.10, respectively) proliferation of endothelial cells compared with control (unconditioned) media. Most of the mitogenic activity of a sample of Fb CM and a sample of Ep CM from one individual uterus bound to heparin-agarose, and each exhibited two major peaks of activity that eluted at 0.9-1.0 and 1.7-1.8 M NaCl; the Fb CM also exhibited an additional heparin-binding peak eluting at 0-0.1 M NaCl. Pooled Fb CM (n = 8 cultures from 7 animals) also contained mitogenic activity for endothelial cells that bound to heparin-agarose, but exhibited three major peaks, eluting at 0.6, 1.1, and 1.8 M NaCl. Pooled Ep CM (n = 4 cultures from 3 animals) showed only one peak of mitogenic activity, which eluted at 0.9 M NaCl. Further characterization indicated that heat treatment reduced the activity of all heparin-binding Fb CM and Ep CM peaks, except the Fb CM peak eluting at 1.7 M NaCl. Trypsin reduced the activity of all peaks except one. Protein-A-purified antibody against fibroblast growth factor 1 (FGF-1) had no or only a slight effect on the mitogenic activity of the peaks. Mitogenic activity of the Fb CM peak eluting at 0.6 M NaCl was reduced by antibody against FGF-2. Activity of the Fb CM and Ep CM peaks that eluted at 1.7-1.8 M NaCl also was immunoneutralized by antibody to FGF-2. These data demonstrate that early passage cultures of endometrial Fb and Ep cells produce heparin-binding endothelial mitogens that appear to be immunologically related to FGF-2. These heparin-binding endothelial mitogens may influence endometrial vascular growth.
Johnson
M L
ML
Department of Animal and Range Sciences, North Dakota State University, Fargo 58105, USA.
Redmer
D A
DA
Reynolds
L P
LP
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000378:metabolism*; D007150:Immunohistochemistry; D008934:Mitogens / Q000378:metabolism*; D014599:Uterus / Q000378:metabolism*
D008934:Mitogens