Organ Culture of Human Main and Accessory Lacrimal Glands and their Secretory Behaviour

The purpose of this study was to investigate the in vitro viability and secretory behaviour of human main and accessory lacrimal glands using an organ culture technique. We evaluated the influence of the second messengers cAMP and cGMP on secretion. Fragments less than 1mm 3of main and accessory lac...

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Veröffentlicht in:Experimental eye research 1996-05, Vol.62 (5), p.541-554
Hauptverfasser: HUNT, S., SPITZNAS, M., SEIFERT, P., RAUWOLF, M.
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SPITZNAS, M.
SEIFERT, P.
RAUWOLF, M.
description The purpose of this study was to investigate the in vitro viability and secretory behaviour of human main and accessory lacrimal glands using an organ culture technique. We evaluated the influence of the second messengers cAMP and cGMP on secretion. Fragments less than 1mm 3of main and accessory lacrimal glands as well as conjunctiva were cultured for 2–72hr at 37°C in an atmosphere consisting of 50% O 2, 45% N 2and 5% CO 2, using a specially devised culture medium (±cAMP or cGMP). The conjunctival tissue served as negative control. Supernatants were assayed for secretory-component-bound IgA, lactoferrin and lysozyme using ELISA. Cultured tissue pieces were embedded in paraffin, serially sectioned, stained and their volumes calculated using an image-analysis system. This enabled us to differentiate between secretory, connective and fatty tissue. Secreted exudate was correlated to the volume of secretory tissue. Viability of cultured organ pieces was determined by electron microscopic examination. Suitable organ culture conditions for human lacrimal glands were successfully established. Electron microscopic examinations proved that the structural characteristics of the organ and the polarity of the individual cells were well preserved up to 22 days of culture. Culture supernatants were assayed for secretory-component-bound IgA, lactoferrin, and lysozyme and showed that the amount of protein secreted increased with time. Upon addition of cAMP (1×10 -3 m) and cGMP (4×10 -3 m), secretion was elevated in both main and accessory lacrimal glands. An organ culture system for lacrimal glands was developed that maintains their structural and cellular characteristics as well as their secretory function for up to 22 days. We believe that this system mimics the in vitro state of the organ better than monolayer cultures and thus proves to be a valuable tool when examining lacrimal function in vitro. The fact that both cAMP and cGMP enhance secretion may help to shed some light on the cellular pathways human main and accessory lacrimal glands use for signal transduction.
doi_str_mv 10.1006/exer.1996.0064
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Electron microscopic examinations proved that the structural characteristics of the organ and the polarity of the individual cells were well preserved up to 22 days of culture. Culture supernatants were assayed for secretory-component-bound IgA, lactoferrin, and lysozyme and showed that the amount of protein secreted increased with time. Upon addition of cAMP (1×10 -3 m) and cGMP (4×10 -3 m), secretion was elevated in both main and accessory lacrimal glands. An organ culture system for lacrimal glands was developed that maintains their structural and cellular characteristics as well as their secretory function for up to 22 days. We believe that this system mimics the in vitro state of the organ better than monolayer cultures and thus proves to be a valuable tool when examining lacrimal function in vitro. The fact that both cAMP and cGMP enhance secretion may help to shed some light on the cellular pathways human main and accessory lacrimal glands use for signal transduction.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>cAMP</subject><subject>cGMP</subject><subject>Cyclic AMP - pharmacology</subject><subject>Cyclic GMP - pharmacology</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Eye and associated structures. Visual pathways and centers. Vision</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>human accessory lacrimal glands</subject><subject>human main lacrimal gland</subject><subject>Humans</subject><subject>Immunoglobulin A, Secretory - analysis</subject><subject>Lacrimal Apparatus - drug effects</subject><subject>Lacrimal Apparatus - secretion</subject><subject>Lacrimal Apparatus - ultrastructure</subject><subject>lactoferrin</subject><subject>Lactoferrin - secretion</subject><subject>lysozyme</subject><subject>Microscopy, Electron</subject><subject>Muramidase - secretion</subject><subject>organ culture</subject><subject>Organ Culture Techniques - methods</subject><subject>Rats</subject><subject>secretory behaviour</subject><subject>secretory-component-bound IgA</subject><subject>Vertebrates: nervous system and sense organs</subject><issn>0014-4835</issn><issn>1096-0007</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kDFPwzAUhC0EKqWwsiFlQGwJtpM4zlgqaJGKOgBitBznmRqlSbGTiv57HBp1Y7L87nvn8yF0TXBEMGb38AM2InnOIn9LTtCY4JyFGOPsFI0xJkmY8Dg9RxfOfflpnGTJCI14luYppWP0sbKfsg5mXdV2FoJGB4tu4wcv0tSBrMtgqhQ419h9sJTKmo2sgnnlBfentmswNngFZaHtmQdYy51pOnuJzrSsHFwN5wS9Pz2-zRbhcjV_nk2XoYoZb8NMS59JAssJLbISONUFjgFLFasUiJacMp0lkjAODCjNMS8YLsokUYziLI8n6O7gu7XNdweuFRvjFFQ-IjSdExn3SyRPPRgdQGUb5yxose1_Y_eCYNE3KfomRd-k6Jv0CzeDc1dsoDziQ3Vevx106ZSstJW1Mu6Ixd6TpsRj_ICBb2Fn_BNOGagVlMaCakXZmP8S_ALmio6v</recordid><startdate>19960501</startdate><enddate>19960501</enddate><creator>HUNT, S.</creator><creator>SPITZNAS, M.</creator><creator>SEIFERT, P.</creator><creator>RAUWOLF, M.</creator><general>Elsevier Ltd</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19960501</creationdate><title>Organ Culture of Human Main and Accessory Lacrimal Glands and their Secretory Behaviour</title><author>HUNT, S. ; SPITZNAS, M. ; SEIFERT, P. ; RAUWOLF, M.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c368t-7fa003ae6912b7de82fb03e0ac3c5e1fa826f74a168e6e22908b60bd44c620793</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>cAMP</topic><topic>cGMP</topic><topic>Cyclic AMP - pharmacology</topic><topic>Cyclic GMP - pharmacology</topic><topic>Enzyme-Linked Immunosorbent Assay</topic><topic>Eye and associated structures. Visual pathways and centers. Vision</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>human accessory lacrimal glands</topic><topic>human main lacrimal gland</topic><topic>Humans</topic><topic>Immunoglobulin A, Secretory - analysis</topic><topic>Lacrimal Apparatus - drug effects</topic><topic>Lacrimal Apparatus - secretion</topic><topic>Lacrimal Apparatus - ultrastructure</topic><topic>lactoferrin</topic><topic>Lactoferrin - secretion</topic><topic>lysozyme</topic><topic>Microscopy, Electron</topic><topic>Muramidase - secretion</topic><topic>organ culture</topic><topic>Organ Culture Techniques - methods</topic><topic>Rats</topic><topic>secretory behaviour</topic><topic>secretory-component-bound IgA</topic><topic>Vertebrates: nervous system and sense organs</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>HUNT, S.</creatorcontrib><creatorcontrib>SPITZNAS, M.</creatorcontrib><creatorcontrib>SEIFERT, P.</creatorcontrib><creatorcontrib>RAUWOLF, M.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Experimental eye research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>HUNT, S.</au><au>SPITZNAS, M.</au><au>SEIFERT, P.</au><au>RAUWOLF, M.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Organ Culture of Human Main and Accessory Lacrimal Glands and their Secretory Behaviour</atitle><jtitle>Experimental eye research</jtitle><addtitle>Exp Eye Res</addtitle><date>1996-05-01</date><risdate>1996</risdate><volume>62</volume><issue>5</issue><spage>541</spage><epage>554</epage><pages>541-554</pages><issn>0014-4835</issn><eissn>1096-0007</eissn><coden>EXERA6</coden><abstract>The purpose of this study was to investigate the in vitro viability and secretory behaviour of human main and accessory lacrimal glands using an organ culture technique. We evaluated the influence of the second messengers cAMP and cGMP on secretion. Fragments less than 1mm 3of main and accessory lacrimal glands as well as conjunctiva were cultured for 2–72hr at 37°C in an atmosphere consisting of 50% O 2, 45% N 2and 5% CO 2, using a specially devised culture medium (±cAMP or cGMP). The conjunctival tissue served as negative control. Supernatants were assayed for secretory-component-bound IgA, lactoferrin and lysozyme using ELISA. Cultured tissue pieces were embedded in paraffin, serially sectioned, stained and their volumes calculated using an image-analysis system. This enabled us to differentiate between secretory, connective and fatty tissue. Secreted exudate was correlated to the volume of secretory tissue. Viability of cultured organ pieces was determined by electron microscopic examination. Suitable organ culture conditions for human lacrimal glands were successfully established. Electron microscopic examinations proved that the structural characteristics of the organ and the polarity of the individual cells were well preserved up to 22 days of culture. Culture supernatants were assayed for secretory-component-bound IgA, lactoferrin, and lysozyme and showed that the amount of protein secreted increased with time. Upon addition of cAMP (1×10 -3 m) and cGMP (4×10 -3 m), secretion was elevated in both main and accessory lacrimal glands. An organ culture system for lacrimal glands was developed that maintains their structural and cellular characteristics as well as their secretory function for up to 22 days. We believe that this system mimics the in vitro state of the organ better than monolayer cultures and thus proves to be a valuable tool when examining lacrimal function in vitro. The fact that both cAMP and cGMP enhance secretion may help to shed some light on the cellular pathways human main and accessory lacrimal glands use for signal transduction.</abstract><cop>London</cop><pub>Elsevier Ltd</pub><pmid>8759522</pmid><doi>10.1006/exer.1996.0064</doi><tpages>14</tpages></addata></record>
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subjects Animals
Biological and medical sciences
cAMP
cGMP
Cyclic AMP - pharmacology
Cyclic GMP - pharmacology
Enzyme-Linked Immunosorbent Assay
Eye and associated structures. Visual pathways and centers. Vision
Fundamental and applied biological sciences. Psychology
human accessory lacrimal glands
human main lacrimal gland
Humans
Immunoglobulin A, Secretory - analysis
Lacrimal Apparatus - drug effects
Lacrimal Apparatus - secretion
Lacrimal Apparatus - ultrastructure
lactoferrin
Lactoferrin - secretion
lysozyme
Microscopy, Electron
Muramidase - secretion
organ culture
Organ Culture Techniques - methods
Rats
secretory behaviour
secretory-component-bound IgA
Vertebrates: nervous system and sense organs
title Organ Culture of Human Main and Accessory Lacrimal Glands and their Secretory Behaviour
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