Name: screening assay for oxidative stress in a feline astrocyte cell line, g355-5
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1. Hücre kültürü ve kedi astrositler H 2 O 2 ile tedavi <ol><li> 37 DMEM medya artı besin ile 75 cm 2 balon Kültür hücre hattı G355-5 ° C ve% 5 CO2,% 60 konfluent kadar (~ 5 gün) . Her 1-2 günde bir medya değiştirin ya da sağlıklı bir kültürü korumak için gerekli. </li><li> Medya çıkarın ve 2 ml% 0.25 tripsin ekleyin. 45 s pipetleme hücreleri Asansör - 1 dakika oda sıcaklığında (RT). </li><li> Süspansiyon, 10 ml medya içeren bir 15 ml konik t?...

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Virüs sıklıkla önerilen bir mekanizma, nöronal hasara oksidatif stres 1, 4, 7, 9, 13, 15, 16, 18-22, 27, 31 neden olmuştur. Temelde, önerilen hidroksit, süperoksit anyon, nitrik oksit ve / veya nöronlar için toksik olan hidrojen peroksit, viral maruz kalma yoluyla glia (astrositler ve mikroglia) serbest reaktif oksijen radikalleri. Benzer şekilde, oksidatif stres de metamfetamin indüklenen nörotoksisite 2, 6, 17 büyük bir patolojik mekanizma olarak öne sürülmüştür. Grubumuzun e...

<table border="1"><tbody><tr><th> Reaktifin Adı </th><th> Şirket </th><th> Katalog numarası </th><th> Yorumlar (isteğe bağlı) </th></tr><tr><td> DMEM </td><td> Cellgro </td><td> 15-013-CV </td><td></td></tr><tr><td> MEM Vitaminler </td><td> Cellgro </td><td> 25-020-cl </td><td> % 1 (v / v) </td></tr><tr><td> L-glutamin </td><td> Hyclone </td><td> 17-605E </td><td> % 1 (v / v) </td></tr><tr><td> Esansiyel Olmayan Amino Asitler </td><td> Hyclone </td><td> 25-025-cl </td><td> % 1 (v / v) </td></tr><tr><td> FBS </td><td> Hyclone </td><td> SH30071.03 </td><td> % 20 (v / v) </td></tr><tr><td> Esansiyel Amino Asitler </td><td> Cellgro </td><td> 25-030-cl </td><td> % 0.2 (v / v) </td></tr><tr><td> 500ml vakum filtrasyon sistemi </td><td> VWR </td><td> 87006-076 </td><td></td></tr><tr><td> 15 ml konik tüpler </td><td> Şahin </td><td> 352097 </td><td></td></tr><tr><td> 75 cm 2 doku kültürü şişeleri </td><td> Şahin </td><td> 353136 </td><td></td></tr><tr><td> 6 sıra doku kültürü plakası </td><td> Şahin </td><td> 353224 </td><td></td></tr><tr><td> CM-H 2 DCFDA </td><td> Invitrogen </td><td> C6827 </td><td></td></tr><tr><td> Propidium İyodür (PI) </td><td> Sigma </td><td> P4170-10mg </td><td></td></tr><tr><td> Tripsin </td><td> Lonza </td><td> 17-160F </td><td></td></tr><tr><td> H 2 O 2 </td><td> Sigma </td><td> H6520 </td><td></td></tr><tr><td> HyQ Antibiyotik </td><td> Hyclone </td><td> SV30079.01 </td><td> % 0.1 (v / v) </td></tr><tr><td> G355-5 hücre </td><td> ATCC </td><td> CRL-2033 </td><td> Normal kedi beyin </td></tr></tbody></table>

screening assay for oxidative stress in a feline astrocyte cell line, g355-5

Nöronal hasara neden bir virüs sık önerilen mekanizma oksidatif stres. Astrositler Santral Sinir Sistemi (MSS) oksidatif stres kontrolünde önemli bir role sahip. Astrositler Reaktif Oksijen Ürünleri (ROS) nöronların yanı sıra korumaya nöronlar için homeostatik çevre korumaya yardımcı olmak. CM-H2DCFDA ROS varlığı için bir hücre geçirgen bir göstergesi. CM-H 2 DCFDA hücre floresan olmayan bir bileşik olarak girer ve hücresel esterazlar asetat grupları kaldırdıktan sonra floresan olur ve bileşik okside . Yeşil floresan olarak bulundu flow sitometri ile ölçülen hücreler, hücreleri oksidatif bir devlet sayısı bir göstergesidir. CM-H 2 DCFDA çok sayıda farklı ROS oksidasyona duyarlıdır. Bu testte ne olursa olsun, hangi oksijen ekrana oksidatif hücresel bir çevre için kullanılacak gibi ROS CM-H 2 DCFDA okside edebilir ilişkin özgüllük Bu eksikliği, bu bileşiğin bir patogenezinde soruşturmasının ilk aşamalarında kullanmak için değerli bir naip yapar ROS radikal ya da kombinasyon hücresel koşulları için sorumludur. Bir kere ROS, CM-H 2 DCFDA oksidasyonu ile mevcut sonra ek deneyler ROS veya Ross birlikte özellikle patogenezinde sürecinde yer aldığı belirlemek için yapılabilir olduğunu kurulmuştur. Bu çalışmanın sonuçları, bu testte G355-5 hücre içinde oksidatif ortamda tespit etmek için değerli bir test, bir olduğunu belirten, hidrojen peroksit CM-H 2 DCFDA bir artış ilavesi ile, tuzlu kontrollere göre tespit edildi floresan olduğunu göstermek kedi astrosit hücre hattı.

Watch this Scientific Journal Video about Screening Assay for Oxidative Stress in a Feline Astrocyte Cell Line, G355-5 at JoVE.com

Screening Assay for Oxidative Stress in a Feline Astrocyte Cell Line, G355-5

Oksidatif hücresel ortamlarda tespit etmek için bir tarama yöntemi, CM-H2DCFDA oksidasyon ölçmek için. Bir kez floresan olmayan bir floresan bileşik bir hücre, CM-H2DCFDA değişiklikler içinde okside. Bu değişiklik, floresan flow sitometri ile ölçüldü ve oksidatif bir ortamda hücrelerin sayısını gösterir.

Feline Astrosit Hücre Hücre Hattı Oksidatif Stres için Tarama Testi, G355-5

An often-suggested mechanism of virus induced neuronal damage is oxidative stress. Astrocytes have an important role in controlling oxidative stress of ...

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Neuroscience

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The Indirect Neuron-astrocyte Coculture Assay: An In Vitro Set-up for the Detailed Investigation of Neuron-glia Interactions

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