MORPHOMETRIC AND FLUORESCENCE ANALYSIS OF NERVE GROWTH FACTOR-DIFFERENTIATED PC12 CELLS

Abstract

Objective PC12 is a rat pheochromocytoma cell line. These cells characteristically undergo differentiation when cultured with nerve growth factor (NGF). Depending on the dose of NGF, the length of neurite extensions changes. Thanks to this differentiation property, the cells are used in neuroscience and in modeling pathophysiological diseases such as Alzheimer's, Parkinson's, and Amyotrophic Lateral Sclerosis. However, literature studies showing the effect of NGF on neurite extensions formed in PC12 cells are very limited. This study aimed to investigate the effect of NGF on neurite extensions and cell viability depending on dose and incubation time. Materials and Methods In this study, PC12 cells were incubated with 50 ng/ml and 100 ng/ml NGF for 3, 6 and 7 days. The lengths of neurite outgrowths and dead cell ratios were calculated in incubated cells. Results The results showed that the length of neurite extensions and dead cell ratio increased depending on NGF doses and incubation time. When NGF incubation times were compared, no difference was found between 50 ng/ml NGF 6 days and 100 ng/ml NGF 3 days groups. Conclusion When the dead cell ratios and sizes of neurite extensions in the experimental groups are evaluated, it is thought that 100 ng/ml NGF and 3 days incubation time parameters are ideal for PC12 cell differentiation.

Keywords

• Nerve growth factor
• Neurite length
• PC12 cell line

SİNİR BÜYÜME FAKTÖRÜ İLE FARKLILAŞTIRILMIŞ PC12 HÜCRELERİNİN MORFOMETRİK VE FLORESANS ANALİZİ

Abstract

Amaç PC12 bir sıçan feokromositoma hücre hattıdır. Bu hücreler, sinir büyüme faktörü (NGF) ile kültürlendiğinde karakteristik olarak farklılaşmaya uğramaktadır. NGF dozuna bağlı olarak, nörit uzantılarının uzunlukları değişir. Bu farklılaşma özelliği sayesinde hücreler nörobilimde ve Alzheimer, Parkinson, Amyotrofik Lateral Skleroz gibi patofizyolojik hastalıkların modellenmesinde kullanılmaktadır. Ancak NGF’nin PC12 hücrelerinde oluşan nörit uzantıları üzerine olan etkisini gösteren literatür çalışmaları oldukça kısıtlıdır. Bu çalışmanın amacı NGF’nin doza ve inkübasyon süresine bağlı olarak nörit uzantıları ve hücre canlılığı üzerine olan etkisini araştırmaktır. Gereç ve Yöntem Bu çalışmada PC12 hücreleri 50 ng/ml ve 100 ng/ml NGF ile 3, 6 ve 7 gün inkübe edilmiştir. İnkübe edilen hücrelerde nörit büyümelerinin uzunlukları ve ölü hücre oranları hesaplanmıştır. Bulgular Elde edilen bulgular ile NGF dozlarına ve inkübasyon süresine bağlı olarak nörit uzantılarının uzunluğunun ve ölü hücre oranının arttığı gösterilmiştir. NGF inkübasyon süreleri karşılaştırıldığında 50 ng/ml NGF 6 gün ve 100 ng/ml NGF 3 gün grupları arasında fark olmadığı bulunmuştur. Sonuç Deney gruplarında ölü hücre oranları ve nörit uzantılarının boyutları değerlendirildiğinde 100 ng/ml NGF ve 3 gün inkübasyon süresi parametrelerinin PC12 hücre farklılaşması için ideal olduğu düşünülmektedir.

Keywords

• Nörit uzunluğu
• PC12 hücre hattı
• Sinir büyüme faktörü

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