Name: on-chip endothelial inflammatory phenotyping
Uploaded: 2012-07-21T18:00:00
Description: Watch this Scientific Journal Video about On-Chip Endothelial Inflammatory Phenotyping at JoVE.com

Bu çalışma Scott I. Simon ve Anthony G. Passerini NIH / NHLBI hibe R01 HL082689 tarafından desteklenmiştir.

1. Hücre Kültürü ve Yüzey Hazırlama <ol><li> Bir torna kullanarak 100 x 20 mm doku kültür kaplarına (BD Falcon) 3-inç dairesel yüzeylerde kesin. % 70 etanol içinde boğulma yolu ile yüzeyler sterilize edin. Oda sıcaklığında 1 saat için 4 ml tip I kollajen (100 ug / ml) içeren bir Petri kabı ve kat yeri, daha sonra 4 ml 1 x PBS ile yıkayın. </li><li> Doğrudan malzeme 1 ml uygulayarak 6.5x10 5 hücre / ml ve tohum İnsan aort endotel hücreleri (HAEC, pasaj 4-6) Askıya. 37 ° C,% 5 <su...

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Biz CAM ifade ve monosit adezyon gerçek zamanlı görüntüleme yoluyla endotelyal inflamatuvar fenotip on-chip değerlendirme için mikroakışkan PDMS cihazların kullanımını açıklar. Bizim yaklaşımın önemli bir avantajı gibi aterojenik damarlarda makaslama stresi ayna tanımlanan hidrodinamik koşullar altında diyet lipid ve sitokinler gibi inflamatuvar mediatörlerin maruz hücrelerde endotel disfonksiyonu ile ilişkili sonuçları sayısal yeteneği yatıyor. VMMC reaktifler veya sınırlı kaynağı ol...

<table border="1"><tbody><tr><td> Madde </td><td> Şirket </td><td> Katalog Numarası </td></tr><tr><td> 100 x 20mm Petri Yemekleri </td><td> BD Falcon </td><td> 353003 </td></tr><tr><td> Etanol,% 95 </td><td> Merck Kimyasallar </td><td> EX0290-1 </td></tr><tr><td> DPBS </td><td> Cellgro </td><td> 21-031-CV </td></tr><tr><td> Ben Sıçan Kuyruğu Türetilmiş Kollajen yazın </td><td> Gibco </td><td> A10483-01 </td></tr><tr><td> İnsan aort endotel hücreleri </td><td> Genlantis </td><td> PH30405A </td></tr><tr><td> Antibiyotik-Antimikotik Çözümü </td><td> Invitrogen </td><td> 15240-062 </td></tr><tr><td> Endotel BulletKit </td><td> Lonza </td><td> CC-4176 </td></tr><tr><td> Endotel Bazal Medya-2 </td><td> Lonza </td><td> CC-3156 </td></tr><tr><td> 10 ml şırıngalar </td><td> BD Falcon </td><td> 309604 </td></tr><tr><td> Poliüretan boru </td><td> Tygon </td><td> ABW0001 </td></tr><tr><td> Leibovitz-15 Medya </td><td> Gibco </td><td> 11415-069 </td></tr><tr><td> Sylgard 184 Silikon Elastomer Base </td><td> Dow Corning </td><td> 184 </td></tr><tr><td> Sylgard 184 Silikon Elastomer Kürleme </td><td> Dow Corning </td><td> 184 </td></tr><tr><td> SU8 fotorezist Usta Gofret </td><td> UC Davis Pan Lab </td><td> N / A </td></tr><tr><td> Eclipse TE200 Ters Mikroskop </td><td> Nikon </td><td> Eclipse TE200 </td></tr><tr><td> Şırınga Pompası </td><td> Harvard Apparatus </td><td> PHD2000 </td></tr><tr><td> 19 numara derialtı iğne </td><td> Kendall </td><td> 8881 </td></tr><tr><td> THP-1 Monositik Hücre Hattı </td><td> ATCC </td><td> TİB-202 </td></tr><tr><td> HBSS Ca 2 + / Mg 2 (Hanks Tuzlu Çözüm Buffered) + </td><td&gt; Gibco </td><td> 14025-092 </td></tr><tr><td> Tümör Nekroz Faktör Alfa (TNF-α) </td><td> R &amp; D Systems </td><td> 210-TA-010 </td></tr><tr><td> Stromal Türemiş Faktörü - 1 (SDF-1) </td><td> R &amp; D Systems </td><td> 350-NS-010 </td></tr><tr><td> RPMI 1640 </td><td> Cellgro </td><td> 10-040-CV </td></tr><tr><td> Albumin Serum İnsan (HSA) </td><td> ZLB Behring </td><td> NDC 0053-7680-32 </td></tr></tbody></table> Tablo 2. Özel reaktifleri ve ekipmanlar.

on-chip endothelial inflammatory phenotyping

Aterogenezin endotelyal disfonksiyona yol açan sistemik enflamasyon artan devlet katkıda metabolik anormallikleri tarafından daha da güçlendirilmektedir. Ancak, risk bireyin düzeyi anlatıyor endotel erken fonksiyonel değişiklikler doğrudan kılavuzu tedavi stratejisi yardım klinik değerlendirilmemektedir. Diğer taraftan, yerel hemodinami ile inflamasyon düzenleme, ateroskleroz, tesadüfi olmayan bir uzaysal dağılımının katkıda bulunur, ama mekanizmalar, in vivo tarif etmek zordur. Biz hassas akış şartlarında insan endotel hücreleri (EC) ve monositler inflamatuar olayların kantitatif tayini metabolik pertürbasyon için bir laboratuvar-on-a-chip bazlı yaklaşım açıklanmaktadır. Yumuşak litografi standart yöntemlerle kültive EC mono tabakalara direkt olarak bağlı olan damar taklit eden microfluidic odaları (VMMC), microfabricate etmek için kullanılır. 1. Bu cihazlar, reaktiflerin küçük miktarlar ise kullanma avantajına sahiptiriyi tanımlanmış bir kesme alanına maruz AK membran inflamatuar olayların doğrudan görüntüleme için bir platform sağlar. Biz başarıyla insan aort EC (HAEC) sitokin-2 lipid-3, 4 ve RAGE indüklenen 5 inflamasyon araştırmak için bu cihazlar uyguladık. Burada tahlil monositik hücre (THP-1) haddeleme ve diferansiyel kayma özellikleri altında klimalıdır ve inflamatuar sitokin TNF-α tarafından aktive edilir HAEC mono tabakaları üzerinde durmasına VMMC kullanımı belgelemek. Bu gibi çalışmalar metabolik risk faktörleri altında atherosusceptibility içine mekanistik anlayış sağlıyor.

Watch this Scientific Journal Video about On-Chip Endothelial Inflammatory Phenotyping at JoVE.com

On-Chip Endothelial Inflammatory Phenotyping

Fotolitografi tarafından kazınmış ve PDMS imal Mikroakışkan akışı odaları EC disfonksiyonu ve inflamasyon ile ilişkili fonksiyonel sonuçlarını araştırmak için uygulanır. Temsili bir deneyde, diferansiyel kayma gerilmesi yeteneği sitokin aktive AK monolayers için monositik hücre adezyon modüle gösterilmiştir.

On-Chip Endotel İnflamatuar fenotipleme

Atherogenesis is potentiated by metabolic abnormalities that contribute to a heightened state of systemic inflammation resulting in endothelial ...

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