Dimetil Sülfoksit, Gliserol ve Metanol’ün HeLa Hücrelerinin Çözdürme Sonrası Hücre Canlılığı Üzerindeki Karşılaştırmalı Etkinliği

Year 2024, Volume: 35 Issue: 1, 32 - 37, 29.03.2024

Zeyno Nuhoğlu Öztürk , Orhan Tokur , Abdurrahman Aksoy

https://doi.org/10.36483/vanvetj.1322291

Abstract

Kriyoprotektanlar, hücrelerin dondurma işlemi sırasında korunması için kullanılır. Bu maddelerin, konsantrasyonu, tipi ve dondurma-çözdürme koşulları kullanılacak hücre tipine göre değişir. Uygun kriyoprezervasyon yönteminin hücreye özel olarak belirlenmesi oldukça önemlidir. Bu çalışma, farklı kriyoprotektanların performansını karşılaştırarak ve bunların çeşitli dondurma ve saklama koşulları altında etkinliğini değerlendirerek, HeLa hücreleri için en uygun kriyoprezervasyon yöntemine ilişkin bilgiler sağlanması amaçlanmaktadır. Hücre süspansiyonları 5:10:85 (v:v:v) oranında kriyoprotektan + fetal sığır serumu + medyumdan oluşan bir dondurucu besiyerinde donduruldu ve 3 ay (-20 °C), 1 ay (-80 °C) ve 6 ay (-80 °C) koşullarında saklandı. Hücre canlılığı ve geri kazanım oranları, çözülmeden hemen sonra ve çözülmeyi takiben 48 saat sonra, tripan mavisi kullanılarak analiz edildi. Canlılık ve geri kazanım oranları 3 ay -20 °C’de, metanol grubunda daha yüksekti. Gliserol grubunda ise canlılık ve geri kazanım oranları 1 ay -80 °C’de daha iyi performans gösterdi. DMSO grubunda ise bu oranlar, 6 ay -80 °C’de en yüksekti. Metanol grubu -80 °C’deki depolama koşullarında başarısız oldu. Bu çalışma, HeLa hücrelerindeki bu kriyoprotektanların, çözdürme işleminden hemen sonra ve 48 saatlik kültivasyondan sonra hücre canlılığı ve hücre geri kazanım oranları üzerindeki etkisini göstermektedir.

Keywords

Dondurarak saklama, Donma, HeLa hücreleri, Hücre kültürü teknikleri, Hücre yaşamı

Comparative Efficacy of the Dimethyl Sulfoxide, Glycerol and Methanol on the Post-Thaw Cell Viability of HeLa Cells

Abstract

Cryoprotectants are used to protect cells during freezing. The concentration, type, and freeze-thaw conditions of these substances vary depending on the type of cell to be used. It is very important to determine the appropriate cryopreservation method for the particular cell. This study aims to provide insights into the optimal cryopreservation method for HeLa cells by comparing the performance of different cryoprotectants and evaluating their effectiveness under various freezing and storage conditions. Cell suspensions were frozen with a freezing media composed of cryoprotectant + fetal bovine serum + medium at a ratio of 5:10:85 (v:v:v) and stored under the following conditions: 3 months (-20 °C), 1 month (-80 °C), and 6 months (-80 °C). Cell viability and recovery rates were analyzed immediately post-thaw and after 48 h using the trypan blue dye exclusion assay. In 3 months (-20 °C), viability and recovery rates were higher in the methanol group. Glycerol showed better performance in 1 month (-80 °C). DMSO was the most efficient in 6 months (-80 °C). Methanol failed at -80 °C storage temperature. This study demonstrates the effect of these cryoprotectants in HeLa cells on cell viability and cell recovery rates immediately after thawing and after 48 hours of cultivation.

Keywords

Cell culture techniques, Cell survival, Cryopreservation, Freezing, HeLa cells

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