Name: preparation of pancreatic acinar cells for the purpose of calcium imaging, cell injury measurements, and adenoviral infection
Uploaded: 2013-07-05T12:00:00
Description: Watch this Scientific Journal Video about Preparation of Pancreatic Acinar Cells for the Purpose of Calcium Imaging, Cell Injury Measurements, and Adenoviral Infection at JoVE.com

Ce travail a été soutenu par un National Institutes of Health Grant DK083327 et DK093491 (à SZH).

1. Préparer les cellules acineuses pancréatiques pour l&#39;imagerie calcique <ol><li> Préparer le tampon d&#39;incubation HEPES contenant HEPES 20 mM, NaCl 95 mM, 4,7 mM de KCl, 0,6 mM MgCl2, 1,3 mM CaCl2, 10 mM de glucose, glutamine 2 mM, et 1 × acides aminés non essentiels moyen de minimum aigle. Ajustez la solution finale à pH 7,4 avec NaOH. </li><li> Préparation d&#39;un tampon d&#39;incubation par addition de BSA (BSA 1% p / v final) à 25 ml du tampon d&#39;incubation HEPES (décrit...

<ol>
	<li>Toescu, E. C., Lawrie, A. M., Petersen, O. H., Gallacher, D. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+and+temporal+distribution+of+agonist-evoked+cytoplasmic+Ca2++signals+in+exocrine+acinar+cells+analysed+by+digital+image+microscopy.">Spatial and temporal distribution of agonist-evoked cytoplasmic Ca2+ signals in exocrine acinar cells analysed by digital image microscopy.</a> Embo J. 11, 1623-1629 (1992).</li><li>Ito, K., Miyashita, Y., Kasai, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Micromolar+and+submicromolar+Ca2++spikes+regulating+distinct+cellular+functions+in+pancreatic+acinar+cells.">Micromolar and submicromolar Ca2+ spikes regulating distinct cellular functions in pancreatic acinar cells.</a> Embo J. 16, 242-251 (1997).</li><li>Husain, S. Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+ryanodine+receptor+mediates+early+zymogen+activation+in+pancreatitis.">The ryanodine receptor mediates early zymogen activation in pancreatitis.</a> Proc. Natl. Acad. Sci. U.S.A. 102, 14386-14391 (2005).</li><li>Raraty, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calcium-dependent+enzyme+activation+and+vacuole+formation+in+the+apical+granular+region+of+pancreatic+acinar+cells.">Calcium-dependent enzyme activation and vacuole formation in the apical granular region of pancreatic acinar cells.</a> Proc. Natl. Acad. Sci. U.S.A. 97, 13126-13131 (2000).</li><li>Orabi, A. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dantrolene+mitigates+caerulein-induced+pancreatitis+in+vivo+in+mice.">Dantrolene mitigates caerulein-induced pancreatitis in vivo in mice.</a> Am. J. Physiol. Gastrointest. Liver Physiol. 299, G196-G204 (2009).</li><li>Shah, A. U., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protease+Activation+during+in+vivo+Pancreatitis+is+Dependent+upon+Calcineurin+Activation.">Protease Activation during in vivo Pancreatitis is Dependent upon Calcineurin Activation.</a> Am. J. Physiol. Gastrointest. Liver Physiol. , (2009).</li><li>Muili, K. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacologic+and+genetic+inhibition+of+calcineurin+protects+against+carbachol-induced+pathologic+zymogen+activation+and+acinar+cell+injury.">Pharmacologic and genetic inhibition of calcineurin protects against carbachol-induced pathologic zymogen activation and acinar cell injury.</a> Am. J. Physiol. Gastrointest. Liver Physiol. , (2012).</li><li>Orabi, A. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ethanol+enhances+carbachol-induced+protease+activation+and+accelerates+Ca2++waves+in+isolated+rat+pancreatic+acini.">Ethanol enhances carbachol-induced protease activation and accelerates Ca2+ waves in isolated rat pancreatic acini.</a> J. Biol. Chem. 286, 14090-14097 (2011).</li><li>Nathanson, M. H., Padfield, P. J., O'Sullivan, A. J., Burgstahler, A. D., Jamieson, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanism+of+Ca2++wave+propagation+in+pancreatic+acinar+cells.">Mechanism of Ca2+ wave propagation in pancreatic acinar cells.</a> J. Biol. Chem. 267, 18118-18121 (1992).</li><li>Reed, A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low+extracellular+pH+induces+damage+in+the+pancreatic+acinar+cell+by+enhancing+calcium+signaling.">Low extracellular pH induces damage in the pancreatic acinar cell by enhancing calcium signaling.</a> J. Biol. Chem. 286, 1919-1926 (2011).</li><li>Amsterdam, A., Jamieson, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structural+and+functional+characterization+of+isolated+pancreatic+exocrine+cells.">Structural and functional characterization of isolated pancreatic exocrine cells.</a> Proc. Natl. Acad. Sci. U.S.A. 69, 3028-3032 (1972).</li><li>Van Acker, G. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+progression+locus-2+is+a+critical+regulator+of+pancreatic+and+lung+inflammation+during+acute+pancreatitis.">Tumor progression locus-2 is a critical regulator of pancreatic and lung inflammation during acute pancreatitis.</a> J. Biol. Chem. 282, 22140-22149 (2007).</li><li>Ito, K., Miyashita, Y., Kasai, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinetic+control+of+multiple+forms+of+Ca(2+)+spikes+by+inositol+trisphosphate+in+pancreatic+acinar+cells.">Kinetic control of multiple forms of Ca(2+) spikes by inositol trisphosphate in pancreatic acinar cells.</a> J. Cell Biol. 146, 405-413 (1999).</li><li>Leite, M. F., Burgstahler, A. D., Nathanson, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ca2++waves+require+sequential+activation+of+inositol+trisphosphate+receptors+and+ryanodine+receptors+in+pancreatic+acini.">Ca2+ waves require sequential activation of inositol trisphosphate receptors and ryanodine receptors in pancreatic acini.</a> Gastroenterology. 122, 415-427 (2002).</li><li>Paredes, R. M., Etzler, J. C., Watts, L. T., Zheng, W., Lechleiter, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+calcium+indicators.">Chemical calcium indicators.</a> Methods. 46, 143-151 (2008).</li><li>Schild, D., Jung, A., Schultens, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Localization+of+calcium+entry+through+calcium+channels+in+olfactory+receptor+neurones+using+a+laser+scanning+microscope+and+the+calcium+indicator+dyes+Fluo-3+and+Fura-Red.">Localization of calcium entry through calcium channels in olfactory receptor neurones using a laser scanning microscope and the calcium indicator dyes Fluo-3 and Fura-Red.</a> Cell Calcium. 15, 341-348 (1994).</li><li>Saluja, A. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Secretagogue-induced+digestive+enzyme+activation+and+cell+injury+in+rat+pancreatic+acini.">Secretagogue-induced digestive enzyme activation and cell injury in rat pancreatic acini.</a> Am. J. Physiol. 276, 835-842 (1999).</li><li>Husain, S. Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ryanodine+receptors+contribute+to+bile+acid-induced+pathological+calcium+signaling+and+pancreatitis+in+mice.">Ryanodine receptors contribute to bile acid-induced pathological calcium signaling and pancreatitis in mice.</a> Am. J. Physiol. Gastrointest. Liver Physiol. , (2012).</li><li>Gurda, G. T., Guo, L., Lee, S. H., Molkentin, J. D., Williams, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cholecystokinin+activates+pancreatic+calcineurin-NFAT+signaling+in+vitro+and+in+vivo.">Cholecystokinin activates pancreatic calcineurin-NFAT signaling in vitro and in vivo.</a> Mol. Biol. Cell. 19, 198-206 (2008).</li></ol>

La méthode d&#39;isolement cellulaire et des essais ultérieurs décrits ici sont des outils puissants pour étudier les caractéristiques physiologiques et physiopathologiques du pancréas exocrine. La méthode pour isoler des cellules acineuses dispersées a été décrite par Amsterdam et Jamieson en 1972 11. Les méthodes présentées ici ont été adaptés à partir de méthodes d&#39;isolement plus récents décrits par Van Acker et ses collègues 12. Bien que ces techniques sont hautement r...

Changements dynamiques de calcium cytosolique sont nécessaires pour les événements de cellules acineuses la fois physiologiques et pathologiques. Ces effets divergents de calcium sont censés résulter de configurations spatiales et temporelles distinctes de la signalisation calcique 1. Par exemple, une enzyme et la sécrétion d&#39;un fluide à partir de la cellule des cellules acineuses sont liés à des pics de calcium à partir d&#39;une région limitée du pôle apical où la sécrétion a lieu 2. En revanche, une vague de calcium global, suivi par des signaux calciques non oscillatoires intense est associé à des événements pathologiques précoces qui conduisent à une pancréatite aiguë 3,4. Il s&#39;agit notamment de l&#39;activation intra-acinaire protéase, la sécrétion de l&#39;enzyme réduite, et la lésion des cellules acineuses. Notre laboratoire utilise isolé acini pancréatiques pour étudier ces événements pathologiques précoces qui conduisent à la maladie à la fois in vivo et in vitro 5-7. Les méthodes décrites ci-dessous décrivent l&#39;isolement des cellules acineuses primaires dans le but de mesurer cyles niveaux de calcium tosolic et lésions cellulaires. Une méthode d&#39;infection par adénovirus de ces cellules est également fournie.

<table border="1">
 <tbody>
 <tr>
 <td>Name</td>
 <td>Company</td>
 <td>Catalogue Number</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>Mice</td>
 <td>NCI</td>
 <td>N/A</td>
 <td>Male 20-30 grams; virtually any strain should yield comparable results.</td>
 </tr>
 <tr>
 <td>HEPES</td>
 <td>American Bioanalytical</td>
 <td>AB00892</td>
 <td></td>
 </tr>
 <tr>
 <td>Sodium Chloride</td>
 <td>J.T. Baker</td>
 <td>3624-05</td>
 <td></td>
 </tr>
 <tr>
 <td>Potassium Chloride</td>
 <td>J.T. Baker</td>
 <td>3040-01</td>
 <td></td>
 </tr>
 <tr>
 <td>Magnesium Chloride</td>
 <td>Sigma</td>
 <td>M-8266</td>
 <td></td>
 </tr>
 <tr>
 <td>Calcium Chloride</td>
 <td>Fischer</td>
 <td>C79</td>
 <td></td>
 </tr>
 <tr>
 <td>Dextrose</td>
 <td>J.T. Baker</td>
 <td>1916-01</td>
 <td></td>
 </tr>
 <tr>
 <td>L-Glutamine</td>
 <td>Sigma</td>
 <td>G-8540</td>
 <td></td>
 </tr>
 <tr>
 <td>1X minimum Eagle's medium non-essential amino acid mixture</td>
 <td>Gibco</td>
 <td>11140-050</td>
 <td></td>
 </tr>
 <tr>
 <td>Sodium Hydroxide</td>
 <td>EM</td>
 <td>SX0593</td>
 <td></td>
 </tr>
 <tr>
 <td>Bovine Serum Albumin</td>
 <td>Sigma</td>
 <td>A7906</td>
 <td></td>
 </tr>
 <tr>
 <td>Collagenase</td>
 <td>Worthington</td>
 <td>4188</td>
 <td></td>
 </tr>
 <tr>
 <td>Soybean Trypsin Inhibitor</td>
 <td>Sigma</td>
 <td>T-9003</td>
 <td></td>
 </tr>
 <tr>
 <td>Carbon Dioxide</td>
 <td>Matheson Gas</td>
 <td> 124-38-9 </td>
 <td></td>
 </tr>
 <tr>
 <td>125 ml Erlenmeyer plastic flask</td>
 <td>Crystalgen </td>
 <td>26-0005</td>
 <td></td>
 </tr>
 <tr>
 <td>Dissection kit</td>
 <td>Fine Science Tools</td>
 <td>14161-10</td>
 <td></td>
 </tr>
 <tr>
 <td>70% Ethanol</td>
 <td>LabChem</td>
 <td>LC222102</td>
 <td></td>
 </tr>
 <tr>
 <td>P1000, P100, P10 pipettes</td>
 <td>Gilson</td>
 <td>FA10005P</td>
 <td></td>
 </tr>
 <tr>
 <td>Weighing boat</td>
 <td>Heathrow Scientific</td>
 <td>HS1420A</td>
 <td></td>
 </tr>
 <tr>
 <td>Plastic transfer pipettes</td>
 <td>USA Scientific</td>
 <td>1020-2500</td>
 <td></td>
 </tr>
 <tr>
 <td>15 ml conical tubes</td>
 <td>BD Falcon</td>
 <td>352095</td>
 <td></td>
 </tr>
 <tr>
 <td>50 ml conical tubes</td>
 <td>BD Falcon</td>
 <td>352070</td>
 <td></td>
 </tr>
 <tr>
 <td>1.5 ml micro-centrifuge tube</td>
 <td>Fisher</td>
 <td>05-408-129</td>
 <td></td>
 </tr>
 <tr>
 <td>0.65 ml micro-centrifuge tube</td>
 <td>VWR</td>
 <td>20170-293</td>
 <td></td>
 </tr>
 <tr>
 <td>22 x 22 mm glass coverslips</td>
 <td>Fisher</td>
 <td>032811-9</td>
 <td></td>
 </tr>
 <tr>
 <td>Nitric Acid</td>
 <td>Fischer</td>
 <td>A483-212</td>
 <td></td>
 </tr>
 <tr>
 <td>Hydrochloric Acid</td>
 <td>Fischer</td>
 <td>A142-212</td>
 <td></td>
 </tr>
 <tr>
 <td>Deionized water</td>
 <td>N/A</td>
 <td>N/A</td>
 <td></td>
 </tr>
 <tr>
 <td>18 x 18 mm coverslips</td>
 <td>Fischer</td>
 <td>021510-9</td>
 <td></td>
 </tr>
 <tr>
 <td>Laboratory film</td>
 <td>Parafilm</td>
 <td>PM-996</td>
 <td></td>
 </tr>
 <tr>
 <td>Fluo-4AM</td>
 <td>Invitrogen</td>
 <td>F14201</td>
 <td></td>
 </tr>
 <tr>
 <td>Dimethylsulfoxide</td>
 <td>Sigma</td>
 <td>D2650</td>
 <td></td>
 </tr>
 <tr>
 <td>Luer lock</td>
 <td>Becton Dickinson</td>
 <td>932777</td>
 <td></td>
 </tr>
 <tr>
 <td>60 ml syringe</td>
 <td>BD Bioscience</td>
 <td>DG567805</td>
 <td></td>
 </tr>
 <tr>
 <td>23 &frac34; gauge needle</td>
 <td>BD Bioscience</td>
 <td>9328270</td>
 <td></td>
 </tr>
 <tr>
 <td>PE50 tubing</td>
 <td>Clay Adam</td>
 <td>PE50-427411</td>
 <td></td>
 </tr>
 <tr>
 <td>Flat head screwdriver </td>
 <td>N/A</td>
 <td>N/A</td>
 <td></td>
 </tr>
 <tr>
 <td>DMEM F-12, no Phenol Red</td>
 <td>Gibco</td>
 <td>21041-025 </td>
 <td></td>
 </tr>
 <tr>
 <td>30.5 gauge needle</td>
 <td>BD Bioscience</td>
 <td>305106</td>
 <td></td>
 </tr>
 <tr>
 <td>5 CC syringe</td>
 <td>BD Bioscience</td>
 <td>309603</td>
 <td></td>
 </tr>
 <tr>
 <td>25 ml Erlenmeyer flask</td>
 <td>Fischer</td>
 <td>FB50025</td>
 <td></td>
 </tr>
 <tr>
 <td>Nylon mesh filter </td>
 <td>Nitex</td>
 <td>03-150/38</td>
 <td>150 &mu;m pore size</td>
 </tr>
 <tr>
 <td>48 well tissue culture plate</td>
 <td>Costar</td>
 <td>3548</td>
 <td></td>
 </tr>
 <tr>
 <td>96 well tissue culture plate </td>
 <td>Costar</td>
 <td>3795</td>
 <td></td>
 </tr>
 <tr>
 <td>6 well tissue culture plate</td>
 <td>Costar</td>
 <td>3506</td>
 <td></td>
 </tr>
 <tr>
 <td>Liquid nitrogen</td>
 <td>Matheson Gas</td>
 <td>7727-37-9</td>
 <td></td>
 </tr>
 <tr>
 <td>Cytotoxicity assay kit</td>
 <td>Promega</td>
 <td>G1782</td>
 <td></td>
 </tr>
 <tr>
 <td>Adeno-GFP</td>
 <td>N/A</td>
 <td>N/A</td>
 <td>Gift from J. Williams</td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Equipment</td>
 </tr>
 <tr>
 <td>Ring stand with clamps</td>
 <td>United Scientific</td>
 <td>SET462</td>
 <td></td>
 </tr>
 <tr>
 <td>Perifusion chamber</td>
 <td>N/A</td>
 <td>N/A</td>
 <td>Designed by S.Z.H and colleagues at Yale University</td>
 </tr>
 <tr>
 <td>Vacuum line</td>
 <td>Manostat</td>
 <td>72-100-000</td>
 <td></td>
 </tr>
 <tr>
 <td>Water bath with shaker</td>
 <td>Precision Scientific</td>
 <td>51220076</td>
 <td></td>
 </tr>
 <tr>
 <td>Confocal microscope</td>
 <td>Zeiss</td>
 <td>LSM 710</td>
 <td></td>
 </tr>
 <tr>
 <td>BioTek Synergy H1 plate reader</td>
 <td>BioTek</td>
 <td>11-120-534</td>
 <td></td>
 </tr>
 <tr>
 <td>Tissue culture hood</td>
 <td>Nuaire</td>
 <td>NU-425-600</td>
 <td></td>
 </tr>
 <tr>
 <td>Tissue culture Incubator</td>
 <td>Thermo</td>
 <td>3110</td>
 <td></td>
 </tr>
 </tbody>
</table>

preparation of pancreatic acinar cells for the purpose of calcium imaging, cell injury measurements, and adenoviral infection

Les cellules acineuses du pancréas est la principale cellule parenchymateuse du pancréas exocrine et joue un rôle primordial dans la sécrétion d&#39;enzymes pancréatiques dans le canal pancréatique. Il est également le site pour l&#39;initiation d&#39;une pancréatite. Nous décrivons ici comment les cellules acineuses sont isolés de signaux intracellulaires de calcium tout le tissu du pancréas et sont mesurées. En outre, nous décrivons les techniques de transfection de ces cellules avec des constructions adénoviraux, et mesurer ensuite la fuite de lactate déshydrogénase, un marqueur de lésion des cellules, dans des conditions qui induisent des lésions des cellules acineuses in vitro. Ces techniques constituent un outil puissant pour caractériser la physiologie des cellules acineuses et la pathologie.

Un exemple de mesures de calcium des cellules acineuses en réponse à des stimuli physiologiques est fourni à la figure 3. cellules acineuses ont été chargés avec le calcium colorant Fluo-4 et perfusé avec l&#39;acétylcholine, un analogue carbachol (CCH; 1 M)) 8. Les cellules ont réagi, sous la forme d&#39;une onde de calcium qui déclenche dans la région apicale et se propage dans la région basolatérale 3,9. tracés représentatifs illustré à la figure 3B</str...

Watch this Scientific Journal Video about Preparation of Pancreatic Acinar Cells for the Purpose of Calcium Imaging, Cell Injury Measurements, and Adenoviral Infection at JoVE.com

Preparation of Pancreatic Acinar Cells for the Purpose of Calcium Imaging, Cell Injury Measurements, and Adenoviral Infection

Nous décrivons une méthode reproductible de préparation de cellules acineuses pancréatiques de souris à partir d&#39;une souris dans le but d&#39;examiner les signaux calciques des cellules acineuses et des lésions cellulaires avec physiologiquement et pathologiquement stimuli pertinents. Une méthode d&#39;infection par adénovirus de ces cellules est également fournie.

Préparation des cellules acineuses pancréatiques à des fins d&#39;imagerie calcique, mesures de blessures cellulaires, et les infections à adénovirus

The pancreatic acinar  cell is the main parenchymal cell of the exocrine pancreas and plays a primary  role in the secretion of pancreatic enzymes into ...

Research

JoVE Journal

Biology

Preparation of Drosophila S2 cells for Light Microscopy

Preparation of Drosophila S2 cells for Light Microscopy

Préparation des Drosophile Les cellules S2 pour la microscopie optique

The ideal experimental system would be cheap and easy to maintain, amenable to a variety of techniques, and would be supported by an extensive literature ...

Recherche

Biologie

Isolation and Culture of Mouse Primary Pancreatic Acinar Cells

Isolement et culture de cellules primaires de souris acineuses du pancréas

This protocol permits rapid isolation (in less than 1 hr) of murine pancreatic acini, making it possible to maintain them in culture for more than one ...

Imaging Cell Membrane Injury and Subcellular Processes Involved in Repair

Imagerie de la membrane cellulaire des blessures et des processus sous-cellulaires impliquées dans la réparation

The ability of injured cells to heal is a fundamental cellular process, but cellular and molecular mechanisms involved in healing injured cells are poorly ...

Live Cell Imaging of Primary Rat Neonatal Cardiomyocytes Following Adenoviral and Lentiviral Transduction Using Confocal Spinning Disk Microscopy

Imagerie de cellules vivantes primaires de rats néonatals cardiomyocytes Suite à adénovirus et Lentivirales transduction confocale à disque rotatif Microscopie

Primary rat neonatal cardiomyocytes are useful in basic in vitro cardiovascular research because they can be easily isolated in large numbers in a single ...

Assessing the Secretory Capacity of Pancreatic Acinar Cells

Évaluation de la capacité sécrétoire des cellules pancréatiques acineuses

Pancreatic acinar cells produce and secrete digestive enzymes. These cells are organized as a cluster which forms and shares a joint lumen. This work ...

Cytosolic Calcium Measurements in Renal Epithelial Cells by Flow Cytometry

Mesures de calcium cytosolique dans les cellules épithéliales rénales par cytométrie en flux

A variety of cellular processes, both physiological and pathophysiological, require or are governed by calcium, including exocytosis, mitochondrial ...

Nephrotoxin Microinjection in Zebrafish to Model Acute Kidney Injury

Néphrotoxine microinjection dans Zebrafish au Modèle lésion rénale aiguë

The kidneys are susceptible to harm from exposure to chemicals they filter from the bloodstream. This can lead to organ injury associated with a rapid ...

Cystometric and External Urethral Sphincter Measurements in Awake Rats with Implanted Catheter and Electrodes Allowing for Repeated Measurements

Mesures de Sphincter urétral cystométriques et externe chez le rat éveillé avec cathéter implanté et électrodes permettant des mesures répétées

Lower urinary tract function is mainly assessed by means of cystometric bladder function analysis in rodents. Conventional cystometries are usually ...

Micromanipulation Techniques Allowing Analysis of Morphogenetic Dynamics and Turnover of Cytoskeletal Regulators

Techniques de micromanipulation permettant l’analyse de la dynamique morphogénétique et chiffre d’affaires des organismes de réglementation du cytosquelette