Dissociated and Explant Cultures of Myenteric Neurons of Pig Small Intestine
Identifieur interne : 000F64 ( Main/Merge ); précédent : 000F63; suivant : 000F65Dissociated and Explant Cultures of Myenteric Neurons of Pig Small Intestine
Auteurs : C. Van Ginneken [Belgique] ; U. Rauch [Allemagne] ; A. Weyns [Belgique] ; K. Sch Fer [Allemagne]Source :
- Anatomia, Histologia, Embryologia [ 0340-2096 ] ; 2005-12.
Abstract
The gastrointestinal tract of pig is a good and available (slaughterhouse material) model to study the enteric nervous system in relation to nutrition, inflammation, development and disease in general. In order to investigate the responses of the enteric nervous network to a variety of stimuli, e.g. growth factors, hormones, extracellular matrix components, but also noxious compounds in a controlled manner, an in vitro experimental set‐up is most appropriate. Methods to obtain in vitro cultures of enteric neurons of rodents, chicken and human are well described. This study attempted to use these methods on pig material. The muscle layer containing the myenteric plexus was dissected from pieces of fetal, neonatal and adult pig jejunum. They were rinsed in Hanks basal salt solution (BSS) in which antibiotics were added. Pieces of ±25 mm2 were transferred to BSS with 1 mg/ml collagenase and 1 mg/ml trypsin inhibitor and incubated for 60 min (5% CO2, 90% RH). Subsequently, they were vortexed for 20 s and ganglia were selected. The procedure was repeated ±4 times. Myenteric ganglia could then be plated or further dissociated in 1 mg/ml trypsin in BSS for 30 min in the incubator. Afterwards, the dissociated ganglia were centrifuged (5 min 1500 rpm) and the trypsin‐solution was replaced with Dulbecco's minimal essential medium (DMEM). The explants or dissociated myenteric ganglia were transferred on non‐coated or coated (poly‐L‐lysine, laminin, fibronectin, collagen or extracellular matrix gel) cover slips. After incubating for 45 min they were topped with 1 ml of DMEM with antibiotics and with or without fetal calf serum (5%). Medium was replaced twice a week. Using immunohistochemistry, both neurons (PGP9.5) and glial cells (S100) could be identified in both culture types. When cultivated under harsh conditions, the dissociated cultures gave rise to neurosphere‐like bodies, containing neurons and glial cells. Thus, the digestion and dissociation technique is applicable to pig material.
Url:
DOI: 10.1111/j.1439-0264.2005.00669_121.x
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: 001683
- to stream Istex, to step Curation: 001681
- to stream Istex, to step Checkpoint: 000829
Links to Exploration step
ISTEX:2B9D4FD4D2BAFF25F4CB6EF44A0D5FD37D487C9ELe document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Dissociated and Explant Cultures of Myenteric Neurons of Pig Small Intestine</title><author><name sortKey="Van Ginneken, C" sort="Van Ginneken, C" uniqKey="Van Ginneken C" first="C." last="Van Ginneken">C. Van Ginneken</name></author><author><name sortKey="Rauch, U" sort="Rauch, U" uniqKey="Rauch U" first="U." last="Rauch">U. Rauch</name></author><author><name sortKey="Weyns, A" sort="Weyns, A" uniqKey="Weyns A" first="A." last="Weyns">A. Weyns</name></author><author><name sortKey="Sch Fer, K" sort="Sch Fer, K" uniqKey="Sch Fer K" first="K." last="Sch Fer">K. Sch Fer</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:2B9D4FD4D2BAFF25F4CB6EF44A0D5FD37D487C9E</idno><date when="2005" year="2005">2005</date><idno type="doi">10.1111/j.1439-0264.2005.00669_121.x</idno><idno type="url">https://api.istex.fr/document/2B9D4FD4D2BAFF25F4CB6EF44A0D5FD37D487C9E/fulltext/pdf</idno><idno type="wicri:Area/Istex/Corpus">001683</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">001683</idno><idno type="wicri:Area/Istex/Curation">001681</idno><idno type="wicri:Area/Istex/Checkpoint">000829</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">000829</idno><idno type="wicri:doubleKey">0340-2096:2005:Van Ginneken C:dissociated:and:explant</idno><idno type="wicri:Area/Main/Merge">000F64</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Dissociated and Explant Cultures of Myenteric Neurons of Pig Small Intestine</title><author><name sortKey="Van Ginneken, C" sort="Van Ginneken, C" uniqKey="Van Ginneken C" first="C." last="Van Ginneken">C. Van Ginneken</name><affiliation wicri:level="4"><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Veterinary Anatomy & Embryology, Department of Veterinary Medicine, Faculty of Farmaceutical, Biomedical and Veterinary Sciences, University of Antwerp, universiteitsplein 1, 2610 Antwerp</wicri:regionArea><orgName type="university">Université d'Anvers</orgName><placeName><settlement type="city">Anvers</settlement><region>Région flamande</region><region type="district" nuts="2">Province d'Anvers</region></placeName></affiliation></author><author><name sortKey="Rauch, U" sort="Rauch, U" uniqKey="Rauch U" first="U." last="Rauch">U. Rauch</name><affiliation wicri:level="1"><country xml:lang="fr">Allemagne</country><wicri:regionArea>Lehrgebiet Biotechnologie, Fachbereich Informatik und Mikrosystemtechnik, Fachhochschule Kaiserslautern, Standort Zweibrücken, Amerikastrasse 1, D‐66482 Zweibrücken</wicri:regionArea><wicri:noRegion>66482 Zweibrücken</wicri:noRegion><wicri:noRegion>D‐66482 Zweibrücken</wicri:noRegion></affiliation></author><author><name sortKey="Weyns, A" sort="Weyns, A" uniqKey="Weyns A" first="A." last="Weyns">A. Weyns</name><affiliation wicri:level="4"><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Veterinary Anatomy & Embryology, Department of Veterinary Medicine, Faculty of Farmaceutical, Biomedical and Veterinary Sciences, University of Antwerp, universiteitsplein 1, 2610 Antwerp</wicri:regionArea><orgName type="university">Université d'Anvers</orgName><placeName><settlement type="city">Anvers</settlement><region>Région flamande</region><region type="district" nuts="2">Province d'Anvers</region></placeName></affiliation></author><author><name sortKey="Sch Fer, K" sort="Sch Fer, K" uniqKey="Sch Fer K" first="K." last="Sch Fer">K. Sch Fer</name><affiliation wicri:level="1"><country xml:lang="fr">Allemagne</country><wicri:regionArea>Lehrgebiet Biotechnologie, Fachbereich Informatik und Mikrosystemtechnik, Fachhochschule Kaiserslautern, Standort Zweibrücken, Amerikastrasse 1, D‐66482 Zweibrücken</wicri:regionArea><wicri:noRegion>66482 Zweibrücken</wicri:noRegion><wicri:noRegion>D‐66482 Zweibrücken</wicri:noRegion></affiliation></author></analytic><monogr></monogr><series><title level="j">Anatomia, Histologia, Embryologia</title><idno type="ISSN">0340-2096</idno><idno type="eISSN">1439-0264</idno><imprint><publisher>Blackwell Verlag GmbH</publisher><pubPlace>Berlin, Germany</pubPlace><date type="published" when="2005-12">2005-12</date><biblScope unit="volume">34</biblScope><biblScope unit="supplement">s1</biblScope><biblScope unit="page" from="53">53</biblScope><biblScope unit="page" to="53">53</biblScope></imprint><idno type="ISSN">0340-2096</idno></series><idno type="istex">2B9D4FD4D2BAFF25F4CB6EF44A0D5FD37D487C9E</idno><idno type="DOI">10.1111/j.1439-0264.2005.00669_121.x</idno><idno type="ArticleID">AHE669_121_121</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0340-2096</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The gastrointestinal tract of pig is a good and available (slaughterhouse material) model to study the enteric nervous system in relation to nutrition, inflammation, development and disease in general. In order to investigate the responses of the enteric nervous network to a variety of stimuli, e.g. growth factors, hormones, extracellular matrix components, but also noxious compounds in a controlled manner, an in vitro experimental set‐up is most appropriate. Methods to obtain in vitro cultures of enteric neurons of rodents, chicken and human are well described. This study attempted to use these methods on pig material. The muscle layer containing the myenteric plexus was dissected from pieces of fetal, neonatal and adult pig jejunum. They were rinsed in Hanks basal salt solution (BSS) in which antibiotics were added. Pieces of ±25 mm2 were transferred to BSS with 1 mg/ml collagenase and 1 mg/ml trypsin inhibitor and incubated for 60 min (5% CO2, 90% RH). Subsequently, they were vortexed for 20 s and ganglia were selected. The procedure was repeated ±4 times. Myenteric ganglia could then be plated or further dissociated in 1 mg/ml trypsin in BSS for 30 min in the incubator. Afterwards, the dissociated ganglia were centrifuged (5 min 1500 rpm) and the trypsin‐solution was replaced with Dulbecco's minimal essential medium (DMEM). The explants or dissociated myenteric ganglia were transferred on non‐coated or coated (poly‐L‐lysine, laminin, fibronectin, collagen or extracellular matrix gel) cover slips. After incubating for 45 min they were topped with 1 ml of DMEM with antibiotics and with or without fetal calf serum (5%). Medium was replaced twice a week. Using immunohistochemistry, both neurons (PGP9.5) and glial cells (S100) could be identified in both culture types. When cultivated under harsh conditions, the dissociated cultures gave rise to neurosphere‐like bodies, containing neurons and glial cells. Thus, the digestion and dissociation technique is applicable to pig material.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Wicri/Eau/explor/EsturgeonV1/Data/Main/Merge
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000F64 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Merge/biblio.hfd -nk 000F64 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Wicri/Eau
|area= EsturgeonV1
|flux= Main
|étape= Merge
|type= RBID
|clé= ISTEX:2B9D4FD4D2BAFF25F4CB6EF44A0D5FD37D487C9E
|texte= Dissociated and Explant Cultures of Myenteric Neurons of Pig Small Intestine
}}
| This area was generated with Dilib version V0.6.27. |  |