Document Detail


Morphology and function of lacrimal gland acinar cells in primary culture.
MedLine Citation:
PMID:  2536359     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The objectives of the current investigation were fourfold: (1) to establish an effective procedure for the isolation of acinar cells from the rat lacrimal gland; (2) to evaluate the functional capacity of freshly isolated cells; (3) to determine defined culture conditions which permit maintenance of viable, differentiated cells, as well as secretory component (SC) production, during long-term culture; and (4) to characterize the morphological features of cultured cells. Acinar cells were isolated by serial incubation of gland fragments in chelating and enzymatic solutions, followed by centrifugation through a Ficoll gradient. The yield of viable cells/gland appeared to be age-dependent: cell recovery was inversely proportional to the age of the animals. Immunofluorescence analysis of freshly isolated cells showed the presence of SC, the IgA antibody receptor, within isolated cells. In addition, experiments with a labeled analog (Nle4-D-Phe7-alpha MSH) of alpha-melanocyte-stimulating hormone (alpha-MSH) demonstrated specific binding sites on freshly isolated cells; alpha-MSH is a known modulator of acinar protein secretion. Maximum binding of the alpha-MSH analog occurred within 30 min, was dependent upon cell density and was reduced by coincubation with unlabeled alpha-MSH. To determine the culture requirements of acinar cells, cells were cultured on a variety of substrates (plastic or modified plastic [Primaria], coated with or without extracellular matrix [Matrigel]) in the presence or absence of various supplements and/or fetal calf serum (FCS) for 0.7 to 3.5 weeks. Cell attachment, function and long-term viability required an extracellular matrix. Moreover, in long term cultures (25 days), acinar cell attachment was enhanced by the inclusion of supplements to media containing 10% FCS. Replacement of serum with fibroblast growth factor, high-density lipoprotein and an increased concentration of epidermal growth factor resulted in a distinct "cobblestone" morphology characteristic of epithelial cell cultures. Electron microscopic analysis of cells cultured in supplemented serum-free media demonstrated extensive rough endoplasmic reticulum and Golgi, intermediate filaments and numerous secretory granules, as well as tight junctions and desmosomes. In addition to cell maintenance and attachment, acinar cell synthesis and/or secretion of SC was positively influenced by inclusion of supplements in the media. In summary, we have isolated lacrimal gland acinar cells, which express receptors for IgA antibodies and alpha-MSH. In addition, we have defined culture conditions which permit the long-term maintenance of SC-secreting acinar cells.
Authors:
L E Hann; J B Tatro; D A Sullivan
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  30     ISSN:  0146-0404     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  1989 Jan 
Date Detail:
Created Date:  1989-02-28     Completed Date:  1989-02-28     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  145-58     Citation Subset:  IM    
Affiliation:
Immunology Unit, Eye Research Institute, Boston, MA 02114.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Adhesion
Cell Separation
Cell Survival
Cells, Cultured
Culture Media
Lacrimal Apparatus / cytology*,  metabolism,  ultrastructure
Male
Microscopy, Electron
Rats
Rats, Inbred Strains
Secretory Component / biosynthesis,  metabolism
Time Factors
alpha-MSH / metabolism
Grant Support
ID/Acronym/Agency:
AM-16684/AM/NIADDK NIH HHS; DK-07927/DK/NIDDK NIH HHS; EY-05612/EY/NEI NIH HHS
Chemical
Reg. No./Substance:
0/Culture Media; 0/Secretory Component; 581-05-5/alpha-MSH

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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