Name: co-culture models of pseudomonas aeruginosa biofilms grown on live human airway cells
Uploaded: 2010-10-06T17:00:00
Description: Watch this Scientific Journal Video about Co-culture Models of Pseudomonas aeruginosa Biofilms Grown on Live Human Airway Cells at JoVE.com

Biz bu modellerin geliştirilmesinde rehberlik ve önerileriniz için G. Ç Toole teşekkür etmek istiyorum. Bu çalışma, Kistik Fibrozis Vakfı (ANDERS06F0 GGA, STANTO07RO ve STANTO08GA BAS), Ulusal Sağlık Enstitüleri (GGA ve R01-HL074175 - BAS T32A107363) tarafından desteklenen ve Biyomedikal Araştırma Mükemmellik için, Ulusal Araştırma Kaynakları Merkezleri Merkezi (BAS Cobre P20-RR018787).

1. Statik Co-kültür Biyofilm Modeli <ol><li> Statik Co-kültür Biyofilm Model 1 aslında ΔF508 CFTR mutasyon 2,3,4 için CF homozigot bireyin geliştirilen hücrelerin ölümsüzleştirdi CFBE41o-hücrelerinin (CFBE hücre), kullanır. CFBE hücreleri, / 6 iyi doku kültürü plakası veya 2 x 10 5% 10 fetal sığır serumu ile desteklenen 24-kuyu asgari bir temel orta doku kültürü plaka (MEM), 10 6 hücreli bir konsantrasyon numaralı seribaşı olmalıdır 2mm L-glu...

<ol>
	<li>Anderson, G. G., Moreau-Marquis, S., Stanton, B. A., O'Toole, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+analysis+of+tobramycin-treated+Pseudomonas+aeruginosa+biofilms+on+cystic+fibrosis-derived+airway+epithelial+cells.">In vitro analysis of tobramycin-treated Pseudomonas aeruginosa biofilms on cystic fibrosis-derived airway epithelial cells.</a> Infect Immun. 76, 1423-1433 (2008).</li><li>Bruscia, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+CF+cell+lines+corrected+at+DeltaF508-CFTR+locus+by+SFHR-mediated+targeting.">Isolation of CF cell lines corrected at DeltaF508-CFTR locus by SFHR-mediated targeting.</a> Gene Ther. 9, 683-685 (2002).</li><li>Cozens, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CFTR+expression+and+chloride+secretion+in+polarized+immortal+human+bronchial+epithelial+cells.">CFTR expression and chloride secretion in polarized immortal human bronchial epithelial cells.</a> Am J Respir Cell Mol Biol. 10, 38-47 (1994).</li><li>Hentchel-Franks, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activation+of+airway+cl-+secretion+in+human+subjects+by+adenosine.">Activation of airway cl- secretion in human subjects by adenosine.</a> Am J Respir Cell Mol Biol. 31, 140-146 (2004).</li><li>Moreau-Marquis, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+DeltaF508-CFTR+mutation+results+in+increased+biofilm+formation+by+Pseudomonas+aeruginosa+by+increasing+iron+availability.">The DeltaF508-CFTR mutation results in increased biofilm formation by Pseudomonas aeruginosa by increasing iron availability.</a> Am J Physiol Lung Cell Mol Physiol. 295, 25-37 (2008).</li><li>Kuchma, S. L., Connolly, J. P., O'Toole, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+three-component+regulatory+system+regulates+biofilm+maturation+and+type+III+secretion+in+Pseudomonas+aeruginosa.">A three-component regulatory system regulates biofilm maturation and type III secretion in Pseudomonas aeruginosa.</a> J Bacteriol. 187, (2005).</li><li>Heydorn, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+reproducibility+in+flow-chamber+biofilms.">Experimental reproducibility in flow-chamber biofilms.</a> Microbiology. 146 (Pt 10), 2409-2415 (2000).</li><li>Heydorn, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+biofilm+structures+by+the+novel+computer+program+COMSTAT.">Quantification of biofilm structures by the novel computer program COMSTAT.</a> Microbiology. 146 (Pt 10), 2395-2407 (2000).</li><li>Anderson, G. G., Yahr, T. L., Lovewell, R. R., O'Toole, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Pseudomonas+aeruginosa+magnesium+transporter+MgtE+inhibits+transcription+of+the+type+III+secretion+system.">The Pseudomonas aeruginosa magnesium transporter MgtE inhibits transcription of the type III secretion system.</a> Infect Immun. 78, (2010).</li><li>Costerton, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Overview+of+microbial+biofilms.">Overview of microbial biofilms.</a> J Ind Microbiol. 15, 137-140 (1995).</li><li>Hoiby, N., Frederiksen, B., Pressler, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eradication+of+early+Pseudomonas+aeruginosa+infection.">Eradication of early Pseudomonas aeruginosa infection.</a> J Cyst Fibros. 4, 49-54 (2005).</li><li>Ko, Y. H., Pedersen, P. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cystic+fibrosis:+a+brief+look+at+some+highlights+of+a+decade+of+research+focused+on+elucidating+and+correcting+the+molecular+basis+of+the+disease.">Cystic fibrosis: a brief look at some highlights of a decade of research focused on elucidating and correcting the molecular basis of the disease.</a> J Bioenerg Biomembr. 33, 513-521 (2001).</li><li>Gibson, R. L., Burns, J. L., Ramsey, B. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathophysiology+and+management+of+pulmonary+infections+in+cystic+fibrosis.">Pathophysiology and management of pulmonary infections in cystic fibrosis.</a> Am J Respir Crit Care Med. 168, 918-951 (2003).</li><li>Furukawa, S., Kuchma, S. L., O'Toole, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Keeping+their+options+open:+acute+versus+persistent+infections.">Keeping their options open: acute versus persistent infections.</a> J Bacteriol. 188, 1211-1217 (2006).</li><li>Merritt, J. H., Kadouri, D. E., O'Toole, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Growing+and+analyzing+static+biofilms.">Growing and analyzing static biofilms.</a> Curr Protoc Microbiol. Chapter 1, Unit 1B 1-Unit 1B 1 (2005).</li><li>O'Toole, G. A., Kolter, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Initiation+of+biofilm+formation+in+Pseudomonas+fluorescens+WCS365+proceeds+via+multiple,+convergent+signalling+pathways:+a+genetic+analysis.">Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways: a genetic analysis.</a> Mol Microbiol. 28, 449-461 (1998).</li><li>Gomez, M. I., Prince, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Opportunistic+infections+in+lung+disease:+Pseudomonas+infections+in+cystic+fibrosis.">Opportunistic infections in lung disease: Pseudomonas infections in cystic fibrosis.</a> Curr Opin Pharmacol. 7, 244-251 (2007).</li></ol>

Biyofilmler çevresel uyaranlara yanıt form bakteri toplulukları. Bu çevresel sinyaller, bir yüzey, toplama, exopolysaccharides üretim ve artan antibiyotik direnci 10 gibi diğer fenotipleri bağlanarak her bakteri içinde küresel düzenleyici değişimlere neden olmaktadır . Son birkaç on yıllar boyunca, pek çok kanıt biyofilmler kronik enfeksiyonların patogenezi büyük bir rol oynadığı hipotezini desteklemiştir. Örneğin, çok iyi kabul edilir P. aeruginosa, biyofilm 11...

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<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>FCS2 (Focht Live-Cell) chamber</td>
<td>&nbsp;</td>
<td>Bioptechs, Butler, PA</td>
<td>060319131616</td>
<td>&nbsp;</td>
<tr>
<td>FCS2 chamber controller</td>
<td>&nbsp;</td>
<td>Bioptechs, Butler, PA</td>
<td>060319-2-0303</td>
<td>&nbsp;</td>
<tr>
<td>40 mm glass coverslips</td>
<td>&nbsp;</td>
<td>Bioptechs, Butler, PA</td>
<td>PH 40-1313-0319</td>
<td>&nbsp;</td>
<tr>
<td>MEM</td>
<td>&nbsp;</td>
<td>Mediatech, Manassass, VA</td>
<td>#10-010-CV</td>
<td>&nbsp;</td>
<tr>
<td>MEM without phenol red</td>
<td>&nbsp;</td>
<td>Mediatech, Manassass, VA</td>
<td>Mediatech, Manassass, VA</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

co-culture models of pseudomonas aeruginosa biofilms grown on live human airway cells

Bakteriyel biyofilm farklı insan hastalıklarının bir numara ile ilişkili olduğu, ancak biyofilm gelişimi genellikle cansız yüzeyler üzerinde çalışılmıştır. Bu çalışmada, kültür ortamında yetişen insan hava yolu epitel hücreleri (CFBE hücre) oluşturan Pseudomonas aeruginosa biyofilm için protokolleri açıklar. İlk yöntem (Statik Co-kültür Biyofilm Modeli olarak adlandırılan), P. aeruginosa, standart doku kültürü plakaları confluent mono tabakaları olarak yetişen CFBE hücreleri ile inkübe edilir . Bakterinin epitel hücreleri oldukça toksik olmasına rağmen, yeterince uzun arginin gecikmeler tek tabaka imha ek biyofilm için CFBE hücreleri oluşturmak için. İkinci yöntem (Akış Hücre Co-kültür Biyofilm Modeli olarak adlandırılır), biyofilm araştırma, CFBE hücrelerinin konfluent tek tabaka destekleyen bir cam lamel karşılamak için genellikle kullanılan bir biyofilm akış hücresi aparatı, adaptasyonu içerir. Bu tek tabaka P. inoküle aeruginosa ve peristaltik bir pompa sonra hücreler boyunca taze orta akar. Her iki sistemde de, bakteriyel biyofilm aşılama sonrası 6-8 saat içinde oluşur. Biyofilm Görselleştirme P. tarafından geliştirilmiş. aeruginosa suşları yapısal yeşil floresan proteininin (GFP) dile getirdi. Statik ve Akış Hücre Co-kültür Biyofilm deneyleri erken P. için model sistemler Kistik Fibrozis (KF) akciğer, ve bu tekniklerin P. aeruginosa enfeksiyonu farklı yönlerini izin aeruginosa biyofilm oluşumu ve virülans biyofilm sitotoksisite, biyofilm CFU ölçümü ve biyofilm boyama ve görselleştirme dahil olmak üzere, ele alınmalıdır.

Watch this Scientific Journal Video about Co-culture Models of Pseudomonas aeruginosa Biofilms Grown on Live Human Airway Cells at JoVE.com

Co-culture Models of Pseudomonas aeruginosa Biofilms Grown on Live Human Airway Cells

Bu kağıt büyüyen farklı yöntemleri açıklar.<em&gt; Pseudomonas aeruginosa</em&gt; Kültürlü insan hava yolu epitel hücreleri üzerinde biyofilm. Bu protokoller, biyofilm birim (CFU) oluşturan koloni ölçme, biyofilm boyama ve biyofilm sitotoksisite eğitimi, biyofilm görselleştirme dahil olmak üzere farklı yönlerini biyofilm oluşumu, çalışma adapte edilebilir.

Co-kültür Modelleri Pseudomonas aeruginosa</emCanlı İnsan Havayolu Hücreler Yetiştirilen> Biyofilmler

Bacterial biofilms have been associated with a number of different human diseases, but biofilm development has generally been studied on non-living ...

Research

JoVE Journal

Immunology and Infection

Co-culture Models of Pseudomonas aeruginosa Biofilms Grown on Live Human Airway Cells

Pseudomonas aeruginosa and Saccharomyces cerevisiae Biofilm in Flow Cells

Pseudomonas aeruginosa and Saccharomyces cerevisiae Biofilm in Flow Cells

Pseudomonas aeruginosa ve Saccharomyces cerevisiae Biyofilm Akış Hücreler

Many microbial cells have the ability to form sessile microbial communities defined as biofilms that have altered physiological and pathological ...

İmmünoloji ve Enfeksiyon

RNA Isolation of Pseudomonas aeruginosa Colonizing the Murine Gastrointestinal Tract

RNA Isolation of Pseudomonas aeruginosa Colonizing the Murine Gastrointestinal Tract

RNA İzolasyonu Pseudomonas aeruginosa Murin Gastrointestinal Yolu koloniler

Pseudomonas aeruginosa (PA) infections result in significant morbidity and mortality in hosts with compromised immune systems, such as patients with ...

Long Term Chronic Pseudomonas aeruginosa Airway Infection in Mice

Long Term Chronic Pseudomonas aeruginosa Airway Infection in Mice

Uzun Vadeli Kronik<em&gt; Pseudomonas aeruginosa</emFarelerde&gt; Havayolu Enfeksiyon

A mouse model of chronic airway infection is a key asset in cystic fibrosis (CF) research, although there are a number of concerns regarding the model ...

Assessing the Innate Sensing of HIV-1 Infected CD4+ T Cells by Plasmacytoid Dendritic Cells Using an Ex vivo Co-culture System.

Assessing the Innate Sensing of HIV-1 Infected CD4+ T Cells by Plasmacytoid Dendritic Cells Using an Ex vivo Co-culture System.

HIV-1 Enfekte CD4 Doğuştan Algılama Değerlendirilmesi<sup&gt; +</sup&gt; Bir kullanılması plazmositoid dendritik hücreler tarafından T Hücreleri<em&gt; Ex vivo</em&gt; Co-kültür sistemi.

HIV-1 innate sensing requires direct contact of infected CD4+ T cells with plasmacytoid dendritic cells (pDCs). In order to study this process, the ...

Pseudomonas aeruginosa Induced Lung Injury Model

Pseudomonas aeruginosa Induced Lung Injury Model

Pseudomonas aeruginosa Kaynaklı Akciğer Hasarı Modeli

In order to study human acute lung injury and pneumonia, it is important to develop animal models to mimic various pathological features of this disease. ...

Replication of the Ordered, Nonredundant Library of Pseudomonas aeruginosa strain PA14 Transposon Insertion Mutants

Replication of the Ordered, Nonredundant Library of Pseudomonas aeruginosa strain PA14 Transposon Insertion Mutants

Sipariş edilmiş, Pseudomonas aeruginosa baskı PA14 Transposon ekleme mutantlar Nonredundant Kütüphane çoğaltılması

Pseudomonas aeruginosa is a phenotypically and genotypically diverse and adaptable Gram-negative bacterium ubiquitous in human environments. P. aeruginosa ...

Methods for Detecting Cytotoxic Amyloids Following Infection of Pulmonary Endothelial Cells by Pseudomonas aeruginosa

Methods for Detecting Cytotoxic Amyloids Following Infection of Pulmonary Endothelial Cells by Pseudomonas aeruginosa

Yöntemleri için algılama sitotoksik Amyloids aşağıdaki enfeksiyon pulmoner endotel hücre Pseudomonas aeruginosa tarafından

Patients who survive pneumonia have elevated death rates in the months following hospital discharge. It has been hypothesized that infection of pulmonary ...

Culture of Small Colony Variant of Pseudomonas aeruginosa and Quantitation of its Alginate

Culture of Small Colony Variant of Pseudomonas aeruginosa and Quantitation of its Alginate

Pseudomonas aeruginosa Küçük Koloni Varyant Kültürü ve Aljinat Quantitation

Pseudomonas aeruginosa, an opportunistic Gram-negative bacterial pathogen, can overproduce an exopolysaccharide alginate resulting in a unique phenotype ...

A Delayed Inoculation Model of Chronic Pseudomonas aeruginosa Wound Infection

A Delayed Inoculation Model of Chronic Pseudomonas aeruginosa Wound Infection

Kronik Pseudomonas aeruginosa Yara Enfeksiyonu Gecikmiş Aşılama Modeli

Pseudomonas aeruginosa (P. aeruginosa) is a major nosocomial pathogen of increasing relevance to human health and disease, particularly in the setting of ...

P. aeruginosa Infected 3D Co-Culture of Bronchial Epithelial Cells and Macrophages at Air-Liquid Interface for Preclinical Evaluation of Anti-Infectives

P. aeruginosa Infected 3D Co-Culture of Bronchial Epithelial Cells and Macrophages at Air-Liquid Interface for Preclinical Evaluation of Anti-Infectives

P. aeruginosa Anti-Enfektiflerin Klinik Öncesi Değerlendirilmesi Için Hava-Sıvı Arabiriminde Bronşiyal Epitel Hücrelerin in enfekte 3D Eş Kültür

fDrug research for the treatment of lung infections is progressing towards predictive in vitro models of high complexity. The multifaceted presence of ...

Co-kültür Modelleri Pseudomonas aeruginosa Biyofilmler