Name: transurethral induction of mouse urinary tract infection
Uploaded: 2010-08-05T17:30:00
Description: Watch this Scientific Journal Video about Transurethral Induction of Mouse Urinary Tract Infection at JoVE.com

De auteurs willen graag Shelly Xie en Jennifer Payne erkennen voor hun deskundige technische bijstand. Dit werk werd mogelijk gemaakt door subsidies van de Society of Pediatric Urologie en de Stanford Pediatric Research Fund.

I. Voorbereiding van de urethrale katheters <ol><li> Voor elke muis blaas en nieren te bemonsteren, Plaats 5-6 roestvrij staal kralen (blaas, 1,6 mm formaat of 0,9-2,0 mm blend) of zirkoniumoxide kralen (nier, 0,5 mm formaat) in een cryovial buis, respectievelijk. Autoclaaf de kraal-bevattende buizen. </li><li> Na de buizen afkoelen tot kamertemperatuur, voeg 200 microliter van steriele PBS aan elk roestvrij staal bead-bevattende buis en 400 microliter van steriele PBS aan elk zirkoonoxide bead-bevattende buis. </li><...

<ol>
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N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+mast+cells+to+bacterial+clearance+and+their+proliferation+during+experimental+cystitis+induced+by+type+1+fimbriated+E.+coli.">Contribution of mast cells to bacterial clearance and their proliferation during experimental cystitis induced by type 1 fimbriated E. coli.</a> Immunol LettM. 91, 103-111 (2004).</li><li>Lane, M. C., Alteri, C. J., Smith, S. N., Mobley, H. 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Inductie van de muis urineweginfectie door transurethrale inoculatie van bacteriën is een reeds lang bestaande, maar technisch veeleisende procedure 11. Hung en Hultgren publiceerde onlangs een uitstekend overzicht van transurethrale technieken van de muis urineweginfectie 13. In dit artikel proberen we verder te illustreren de finesses van deze experimentele aanpak. Beginners kunnen oefenen hun techniek door het uitvoeren van een midline laparotomie om de blaas bloot ...

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		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Cryovial tubes, 2.0 ml polypropylene (no caps)</td>
<td>&nbsp;</td>
<td>E&amp;K Scientific</td>
<td>640201</td>
<td>&nbsp;</td>
<tr>
<td>Caps for cryovial tubes, with 'O' ring</td>
<td>&nbsp;</td>
<td>E&amp;K Scientific</td>
<td>440011-N</td>
<td>&nbsp;</td>
<tr>
<td>Bullet Blender</td>
<td>&nbsp;</td>
<td> Next Advance</td>
<td>BBX24</td>
<td>&nbsp;</td>
<tr>
<td>Zirconium Oxide Beads (0.5mm)</td>
<td>&nbsp;</td>
<td>Next Advance</td>
<td>ZROB05</td>
<td>&nbsp;</td>
<tr>
<td>Stainless Steel Beads (1.6mm)</td>
<td>&nbsp;</td>
<td>Next Advance</td>
<td>SSB16</td>
<td>&nbsp;</td>
<tr>
<td>Stainless Steel Bead Blend (0.9-2.0mm)</td>
<td>&nbsp;</td>
<td>Next Advance</td>
<td>SSB14B</td>
<td>&nbsp;</td>
<tr>
<td>CBA/J Mice, 8-12 week old female</td>
<td>&nbsp;</td>
<td>Jackson Laboratories</td>
<td>000656</td>
<td>&nbsp;</td>
<tr>
<td>Phosphate Buffered Saline (NaCl 137mM, KCl 2.7mM, Na2HPO4 4.3mM, KH2PO4 1.4mM)</td>
<td>&nbsp;</td>
<td>EMD</td>
<td>EM-SX0420-13 (NaCl) EM-PX1405-1 (KCl) EM1.06585.0 (Na2HPO4) EM-5108-1 (KH2PO4)</td>
<td>&nbsp;</td>
<tr>
<td>LB agar</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>LB broth</td>
<td>&nbsp;</td>
<td>EMD</td>
<td>1.10285.0500</td>
<td>&nbsp;</td>
<tr>
<td>Isoflurane (Aerrane)</td>
<td>&nbsp;</td>
<td>Baxter</td>
<td>NDC 10019-773-60</td>
<td>&nbsp;</td>
<tr>
<td>Inoculating loops</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>22-363-601</td>
<td>&nbsp;</td>
<tr>
<td>Eosin-methylene blue (EMB) agar powder</td>
<td>&nbsp;</td>
<td>Himedia Laboratories</td>
<td>MM022-500G</td>
<td>&nbsp;</td>
<tr>
<td>MacConkey agar powder</td>
<td>&nbsp;</td>
<td>Difco</td>
<td>212123</td>
<td>&nbsp;</td>
<tr>
<td>LB agar powder</td>
<td>&nbsp;</td>
<td>Difco</td>
<td>244520</td>
<td>&nbsp;</td>
<tr>
<td>Intramedic non-radiopaque polyethylene tubing, PE10 [inner dimension 0.28 mm (0.011 inch); outer dimension 0.61 mm (0.024 inch)]</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>427401</td>
<td>&nbsp;</td>
<tr>
<td>30 gauge needles, 1/2 inch</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>305106</td>
<td>&nbsp;</td>
<tr>
<td>1 ml tuberculin Slip-Tip syringe</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>309602</td>
<td>&nbsp;</td>
<tr>
<td>3 ml Luer-Lock syringe</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>309585</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

transurethral induction of mouse urinary tract infection

Uropathogene bacteriestammen die van belang zijn gekweekt op agar. In het algemeen, uropathogene<em&gt; E. coli</em&gt; (UPEC) en andere stammen kunnen &#39;s nachts worden geteeld op Luria-Bertani (LB) agar bij 37 ° C in de lucht. UPEC stammen groeien als geel-wit doorschijnende kolonies op LB agar. Na de bevestiging van de juiste kolonie morfologie, zijn enkele kolonies vervolgens opgepikt te worden gekweekt in bouillon. LB bouillon kan gebruikt worden voor de meeste uropathogene bacteriestammen. Twee seriële, &#39;s nachts LB bouillon culturen kan worden gebruikt om de expressie van type I pili, een goed gedefinieerde virulentie factor voor uropathogene bacteriën te verbeteren. Bouillon culturen zijn verdund tot de gewenste concentratie in fosfaat gebufferde zoutoplossing (PBS). Acht tot 12 weken oude vrouwelijke muizen zijn geplaatst onder isofluraan anesthesie en transurethrally geënt met bacteriën met behulp van polyethyleen buizen bedekt met 30 gauge spuiten. Typische inocula, die empirisch moeten worden bepaald voor elke bacteriële / muizenstam combinatie, zijn 10<sup&gt; 6</sup&gt; Tot 10<sup&gt; 8</sup&gt; Kve per muis in 10 tot 50 microliter van PBS. Na de gewenste infectie periode (een dag tot enkele weken), urine monsters en de blaas en de beide nieren worden geoogst. Elk orgaan is gehakt, geplaatst in PBS, en gehomogeniseerd in een Blue Bullet homogenisator. Urine en weefsel homogenaten zijn serieel verdund in PBS en gekweekt op juiste agar. De volgende dag, zijn kolonievormende eenheden geteld.

Watch this Scientific Journal Video about Transurethral Induction of Mouse Urinary Tract Infection at JoVE.com

Transurethral Induction of Mouse Urinary Tract Infection

Deze video zal aantonen methoden om transurethrally veroorzaken muis infecties van de urinewegen en kwantificeren van de omvang van de resulterende infecties.

Transurethrale Inductie van Mouse urineweginfectie

Uropathogenic bacterial strains of interest are grown on agar. Generally, uropathogenic E. coli (UPEC) and other strains can be grown overnight on ...

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