Fetuin: a serum component associated with rat Sertoli and spermatogenic cells in coculture.
Cocultures of rat Sertoli-spermatogenic cells plated in a culture medium supplemented with 10% fetal bovine serum for 6-12 h and then maintained in serum free, hormone/growth factor-supplemented medium accumulated an acidic glycoprotein of molecular weight of 68,000 dalton (68 kD) and isoelectric point range of about 4.2-3.5. Anion exchange chromatography has allowed the partial purification of this protein, which consists of a major protein band of 68 kD and two minor, low molecular weight components. A rabbit antiserum raised against the 68 kD component also crossreacts with the two low molecular weight components, thus suggesting that these two minor components are antigenically related to the 68 kD protein. The 68 kD protein has been identified as fetuin, the major component of fetal bovine serum, based on similar molecular weight, isoelectric point, immunoreactivity and trypsin inhibitory activity. Labeling experiments with [14C]amino acid mixture show that 68 kD protein is not synthesized by cocultured rat Sertoli and spermatogenic cells. Immunocytochemistry and Western blot approaches carried out under various experimental conditions support the view that the fetuin-68 kD protein is taken up from serum by both Sertoli cells and pachytene spermatocytes. Because fetuin 1) behaves as a carrier protein for growth factors, 2) has protease inhibitory activity, 3) is preferentially internalized by Sertoli cells and pachytene spermatocytes and 4) fetal bovine serum-supplemented medium impairs spermatogenic cell viability, there is a need to further define appropriate conditions for optimizing long-term viability and differentiation of spermatogenic cells in vitro.
M
M
Abdullah
J A
JA
Crowell
L L
LL
Tres
A L
AL
Kierszenbaum
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D002852:Chromatography, Ion Exchange; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000032:analysis; D007526:Isoelectric Point; D008297:Male; D008970:Molecular Weight; D051381:Rats; D012708:Sertoli Cells / Q000032:analysis*; D013091:Spermatogenesis; D014357:Trypsin / Q000378:metabolism; D014361:Trypsin Inhibitors / Q000032:analysis; D000509:alpha-Fetoproteins / Q000032:analysis*
D000596:Amino Acids; D006023:Glycoproteins; D014361:Trypsin Inhibitors; D000509:alpha-Fetoproteins; D014357:Trypsin