HT22 hücre hattının farklılaşma karakterinin koloni canlılık testi ile sınanması

Year 2020, Volume: 13 Issue: 3, 331 - 338, 05.12.2020

Nail Can Öztürk

https://doi.org/10.26559/mersinsbd.771704

Cited By: 1

Abstract

Amaç: Bu çalışmanın amacı hali hazırda çeşitli nörodejeneratif hastalıkların modellenmesinde kullanılmakta olan HT22 hücre hattının nörogenez modeli olarak da kullanılabilirliğinin koloni canlılık testi ile sınanmasıdır. Yöntem: Yalnızca HG-DMEM medyumu ile muamele edilen hücreler kontrol grubu (K), 24 saat HG-DMEM medyumda bırakıldıktan sonra sırasıyla 24, 48 ve 72 saat B27+ katkılı NB+ medyumu ile inkübe edilen hücreler ise grup 1 (G1), grup 2 (G2) ve grup 3 (G3) olarak belirlenmiştir. Tüm gruplar daha sonra %0.5’lik kristal violet ile boyanarak görüntülenmiştir. Elde edilen görüntülerden koloni sayımı yapılarak hücre canlılık oranları tayin edilmiş ve istatistiksel analiz gerçekleştirilmiştir. Bulgular: Tüm gruplar arasındaki koloni canlılık oranları açısından istatistiksel olarak anlamlı bir farklılık saptanmıştır (p<0.0001). Farklılığın yönü, kontrol grubuna kıyasla deney gruplarındaki düşüş eğilimi olmakla birlikte; grup 1 ile grup 2 ve grup 3 birbirleriyle karşılaştırıldığında gözlenen azalmanın istatistiksel olarak anlamlı olduğu görülmüştür (p<0.0001). Ancak grup 2 ve grup 3 arasında farklılık görülmemiştir (p=0.254). Sonuç: Özetle, HT22 hücre hattının nörogenez modeli olarak kullanılabilirliği basit ve ucuz bir yöntem ile sınırlı düzeyde olsa da sınanmıştır. Bu çalışmada uygulanan farklılaştırma protokolü kullanılarak hücresel farklılaşmanın değişik basamaklarını temsil eden faktörler gen ve/veya protein ekspresyonu seviyesinde de test edilebilirse, HT22 hücrelerinin yaygın biçimde nörogenez modeli olarak kullanılabilirliği ortaya çıkarılabilir.
Anahtar kelimeler: HT22, hipokampus, nörogenez, nöronal farklılaşma, koloni canlılık testi

Keywords

HT22, Hipokampus, Nörogenez, Nöronal Farklılaşma, Koloni Canlılık Testi

Supporting Institution

Mersin Üniversitesi

Project Number

2018-1-AP4-2875

Thanks

Bu araştırma Mersin Üniversitesi 2018-1-AP4-2875 numaralı BAP projesi tarafından desteklenmiştir. HT22 hücre hattı Marmara Üniversitesi, Biyokimya A.D. öğretim üyesi, Prof. Dr. Betül Karademir tarafından, Mersin Üniversitesi Tıp Fakültesi, Anatomi Anabilim Dalı’na hediye olarak verilmiştir.

Testing the differentiation character of HT22 cell line via colony survival assay

Abstract

Aim: The aim of this study is to test the feasibility of HT22 cell line, which is in a current use in the modeling of various neurodegenerative diseases, as a neurogenesis model by colony survival assay. Method: Group of cells treated with HG-DMEM medium was designated as control group (K) while equivalent counterparts upon the same treatment that subjected to NB + medium supplemented with B27+ for 24 (G1), 48 (G2), and 72 hours (G3), respectively. All groups were then imaged by staining with 0.5% crystal violet under inverted microscope. Cell viability rates were determined by counting the colonies from the visuals obtained and statistical analysis was performed. Results: A statistically significant difference in colony viability rates between different groups was evident (p<0.0001). The direction of difference was in a decrement trend in the experimental groups compared to the control group (p<0.0001). However, when the two individual experimental groups (G2 vs G3) were compared, there was no significance (p=0.254). Conclusion: In summary, feasibility the use of HT22 cell line as a neurogenesis model has been tested in the current study, albeit limited, by a simple and inexpensive method. If the factors representing the different stages of cellular differentiation can also be tested at the gene and / or protein expression level using the differentiation protocol used in this study, the availability of HT22 cells as a neurogenesis model will be better revealed.

Keywords

HT22, hippocampus, neurogenesis, neuronal differentiation, colony survival assay

Project Number

2018-1-AP4-2875

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