The production and properties of a tissue plasminogen activator from normal epithelial cells grown in microcarrier culture.
A plasminogen activator with different biochemical and physical properties to the one obtained from Bowes melanoma cells has been isolated from a normal epithelial cell line derived from guinea-pig keratocytes (GPK). Cell growth and enzyme production is carried out in microcarrier cultures using up to 15 g Cytodex 3 per litre. Cells are maintained in optimum conditions by the use of a closed perfusion loop and the system has been scaled up to 20 litres. Initially enzyme production was in serum-free medium following a growth phase in serum supplemented medium. However, the discovery that the cells produce the enzyme mainly during the replicative phase has led to investigating the use of serum-substitutes and growth factors on cell growth and enzyme production. The aim is to harvest the enzyme after the growth phase which precludes the use of high (greater than 1%) serum concentrations. The biochemical and biophysical properties of the enzyme are described.
D000818:Animals; D002455:Cell Division; D002460:Cell Line; D003315:Cornea / Q000166:cytology / Q000378:metabolism; D003470:Culture Media; D003584:Cytological Techniques; D004848:Epithelium / Q000378:metabolism; D006168:Guinea Pigs; D008863:Microspheres; D010960:Plasminogen Activators / Q000096:biosynthesis*
D003470:Culture Media; D010960:Plasminogen Activators