Influence of culture conditions on the androgen control of secretory component production by acinar cells from the rat lacrimal gland.
Research has shown that androgens regulate the production of secretory component (SC), the IgA antibody receptor, by lacrimal gland acinar cells in vivo. This study was designed to establish an optimal culture system to permit analysis of this endocrine-acinar cell interrelationship in vitro. Acinar cells were isolated from male rat lacrimal glands and cultured on Matrigel (Collaborative Research, Bedford, MA) in serum-free Dulbecco's modified Eagle's medium (DMEM)/Ham's F12 media that contained a variety of supplements. Under these conditions, acinar cells responded to dihydrotestosterone (DHT) exposure with a significant increase in SC output. Replacement of the DMEM/Ham's F12 media base with either Modified Eagle's Medium (MEM) or low-calcium MEM inhibited this hormone response and dramatically reduced cell recovery after 4 days of culture. Similarly, decreased concentrations or deletions of selected media supplements, including insulin, which binds to acinar cells, and dexamethasone, led to a significant diminution in the extent of androgen action, as well as to a decline in cell maintenance. In contrast, removal of high-density lipoprotein from culture media or the addition of fetal bovine serum (FBS) or cholera toxin significantly enhanced basal and DHT-associated SC production by acinar cells. With regard to extracellular matrices, Matrigel proved to be superior to collagen type I, laminin, fibronectin, or the Primaria (Falcon, Oxnard, CA) plastic surface in providing support for acinar cell association or hormone-related function. In summary, our results show that the media formulation, supplement profile, and extracellular matrix composition are important for maximal expression of androgen-induced effects by lacrimal gland acinar cells in vitro.
Hann
L E
LE
Immunology Unit, Eye Research Institute, Boston, MA 02114.
Kelleher
R S
RS
Sullivan
D A
DA
D000728:Androgens / Q000502:physiology*; D000818:Animals; D001665:Binding Sites; D001672:Biocompatible Materials / Q000494:pharmacology; D001790:Blood Physiological Phenomena; D001834:Body Water / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D003584:Cytological Techniques; D007328:Insulin / Q000378:metabolism / Q000494:pharmacology; D007765:Lacrimal Apparatus / Q000166:cytology / Q000378:metabolism*; D008297:Male; D008934:Mitogens / Q000494:pharmacology; D009994:Osmolar Concentration; D051381:Rats; D012635:Secretory Component / Q000096:biosynthesis*; D014168:Transferrin / Q000494:pharmacology
D000728:Androgens; D001672:Biocompatible Materials; D003470:Culture Media; D007328:Insulin; D008934:Mitogens; D012635:Secretory Component; D014168:Transferrin