Serum-free culture of AtT 20 pituitary cells: a system for neuroendocrine studies under defined conditions.
The growth of the mouse pituitary cell line AtT 20 was studied under different in vitro conditions. A completely defined, serum-free culture medium supported the survival of cells for a period of more than 2 mo. The medium, designed SFI, consisted of basal medium supplemented with transferrin, insulin, putrescine, and selenium. For maintenance of cells during long-term culture, no additional compounds were necessary. The time-dependent increases in cell number during culture with fetal bovine serum (FBS) and under serum-free conditions showed similar properties. Analysis of the effects of different substrata on cell growth demonstrated that polylysine supported adhesion and initial growth of cells to a greater extent than untreated plastic or FBS adsorbed to culture dishes. Synthesis and regulation of proopiomelanocortin (POMC)-mRNA, the precursor-mRNA of adrenocorticotropin (ACTH), could be detected by Northern blot analysis under basal conditions and after incubation with steroids and corticotropin-releasing hormone (CRH), indicating the serum-independent expression of important cellular properties.
Vedder
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University of Freiburg, Department of Psychiatry, Federal Republic of Germany.
D000324:Adrenocorticotropic Hormone / Q000235:genetics / Q000378:metabolism; D000818:Animals; D015152:Blotting, Northern; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D003346:Corticotropin-Releasing Hormone / Q000494:pharmacology; D003470:Culture Media / Q000032:analysis / Q000494:pharmacology; D015870:Gene Expression; D007328:Insulin / Q000032:analysis / Q000494:pharmacology; D051379:Mice; D010902:Pituitary Gland / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D011107:Polylysine / Q000494:pharmacology; D011333:Pro-Opiomelanocortin / Q000235:genetics / Q000378:metabolism; D011700:Putrescine / Q000032:analysis / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012643:Selenium / Q000032:analysis / Q000494:pharmacology; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000494:pharmacology; D013997:Time Factors; D014168:Transferrin / Q000032:analysis / Q000494:pharmacology
D003470:Culture Media; D007328:Insulin; D012333:RNA, Messenger; D014168:Transferrin; D011107:Polylysine; D012710:Serum Albumin, Bovine; D011333:Pro-Opiomelanocortin; D000324:Adrenocorticotropic Hormone; D003346:Corticotropin-Releasing Hormone; D012643:Selenium; D011700:Putrescine