The Journal of Faculty of Pharmacy of Ankara University is an international medium, an open access, peer-reviewed journal for the publication of original research reports, reviews and short communications in English or Turkish on relevant developments in pharmaceutical sciences., Proceeding of scientific meetings may be published as special issues of supplements to the journal. Additionally, it contains previous and forthcoming national and international scientific meetings with social activities in pharmaceutical areas.
Research Article
Year 2025,
Volume: 49 Issue: 2, 19 - 19
Abstract
Objective: The effect of storage conditions on isolated lymphocyte viability is a critical factor for the long-term preservation of cells. Lymphocytes are essential cells of the immune system and are often isolated and stored in laboratory environments for different research topics such as chronic diseases, including cancer. This storage process must be conducted under suitable conditions to preserve the cells' physiological and biochemical properties. In this context, determining the effects of storage conditions on the viability of isolated lymphocytes is crucial for obtaining accurate results in research.
Material and Method: This research investigates whether storage conditions affect DNA damage in human lymphocyte samples. Whole blood and isolated lymphocytes were stored at two different temperatures (-20°C and -80°C) for 1, 3, 6, and 12 months. Additionally, for each time point, some of the lymphocytes stored under different temperature conditions were preserved in a cryopreservation solution containing 10% DMSO, while others were stored directly. The aim of this study is to contribute to various research areas, such as human biomonitoring studies, where lymphocytes are used, by evaluating lymphocyte viability under different storage conditions and time periods.
Result and Discussion: According to the findings of our study, the survival rate of lymphocytes was higher under storage conditions at -80°C. Additionally, it can be stated that storing lymphocytes at -80°C with a cryopreservation solution (10% DMSO) provides a more suitable strategy for preserving lymphocyte integrity.
Keywords
References
- 1. Ho, C.K., Choi, S.W., Siu, P.M., Benzie, I.F. (2011). Cryopreservation and storage effects on cell numbers and DNA damage in human lymphocytes. Biopreservation and Biobanking, 9(4), 343-347. [CrossRef]
- 2. Germann, A., Oh, Y.J., Schmidt, T., Schön, U., Zimmermann, H., von Briesen, H. (2013). Temperature fluctuations during deep temperature cryopreservation reduce PBMC recovery, viability and T-cell function. Cryobiology, 67(2), 193-200. [CrossRef]
- 3. Belloni, P., Meschini, R., Palitti, F. (2008). Effects of storage conditions of human whole blood on the viability of lymphocytes. International Journal of Radiation Biology, 84(7), 613-619. [CrossRef]
- 4. Allione, A., Porcedda, P., Russo, A., Ricceri, F., Simonelli, V., Minoprio, A., Guarrera, S., Pardini, B., Mazzei, F., Dogliotti, E., Giachino, C., Matullo, G. (2013). Effect of blood storage conditions on DNA repair capacity measurements in peripheral blood mononuclear cells. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, 749(1-2), 73-79. [CrossRef]
- 5. Jalla, S., Sazawal, S., Deb, S., Black, R. E., Das, S.N., Sarkar, A., Bhan, M.K. (2004). Enumeration of lymphocyte subsets using flow cytometry: Effect of storage before and after staining in a developing country setting. Indian Journal of Clinical Biochemistry, 19, 95-99. [CrossRef]
- 6. Duthie, S.J., Pirie, L., Jenkinson, A.M., Narayanan, S. (2002). Cryopreserved versus freshly isolated lymphocytes in human biomonitoring: Endogenous and induced DNA damage, antioxidant status and repair capability. Mutagenesis, 17(3), 211-214. [CrossRef]
- 7. Belloni, P., Pepe, G., Palitti, F. (2010). Effect of storage conditions of blood on radiation-induced chromosomal aberrations and apoptosis in human lymphocytes. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 695(1-2), 35-39. [CrossRef]
- 8. Park, M.S., Terasaki, P.I. (1974). Storage of human lymphocytes at room temperature. Transplantation, 18(6), 520-524. [CrossRef]
- 9. Stopford, C.R., Macqueen, J.M., Amos, D.B., Ward, F.E. (1972). Some variations in lymphocyte freezing methods which do not affect cell viability. Tissue Antigens, 2(1), 20-26. [CrossRef]
- 10. Vaught, J.B. (2006). Blood collection, shipment, processing, and storage. Cancer Epidemiology Biomarkers & Prevention, 15(9), 1582-1584. [CrossRef]
- 11. Ladeira, C., Koppen, G., Scavone, F., Giovannelli, L. (2019). The comet assay for human biomonitoring: effect of cryopreservation on DNA damage in different blood cell preparations. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 843, 11-17. [CrossRef]
- 12. Ekong, T., Kupek, E., Hill, A., Clark, C., Davies, A., Pinching, A. (1993). Technical influences on immunophenotyping by flow cytometry the effect of time and temperature of storage on the viability of lymphocyte subsets. Journal of Immunological Methods, 164(2), 263-273. [CrossRef]
Year 2025,
Volume: 49 Issue: 2, 19 - 19
Abstract
Amaç: Saklama koşullarının izole lenfosit canlılığı üzerindeki etkisi, hücrelerin uzun süreli saklanabilmesi için kritik bir faktördür. Lenfositler, bağışıklık sisteminin önemli hücreleri olup, çoğunlukla kanser gibi kronik hastalıkların değerlendirilmesi veya farklı araştırma konularında kullanılmak üzere laboratuvar ortamında izole edilip saklanırlar. Bu saklama işlemi, hücrelerin fizyolojik ve biyokimyasal özelliklerini koruyabilmesi için uygun koşullarda yapılmalıdır.
Bu bağlamda, saklama koşullarının izole lenfositlerin canlılığı üzerindeki etkileri belirlemek, yapılacak araştırmalarda doğru sonuç alabilmek için çok önemlidir.
Gereç ve Yöntem: Bu makalede, insan lenfosit örneklerinde saklama koşullarının DNA hasarını etkileyip etkilemediği araştırılmıştır. Tam kan ve İzole lenfositler 1, 3, 6 ve 12 aylık süreler boyunca 2 farklı sıcaklık (-20°C ve -80°C) altında saklanmıştır. Ayrıca, her bir zaman aralığı için farklı sıcaklık koşullarında saklanan lenfositlerin bir kısmı %10 DMSO içeren kriyoprezervasyon çözeltisinde, diğer bir kısmı ise doğrudan saklanmıştır. Bu çalışmayla, farklı sürelerde ve farklı saklama koşulları altında lenfosit canlılıklarını değerlendirerek insan biyoizleme çalışmaları gibi lenfositlerin kullanıldığı farklı çalışma alanlarına elde ettiğimiz veriler ile katkıda bulunmak amaçlanmıştır.
Sonuç ve Tartışma: Çalışmamızda elde edilen bulgulara göre lenfositlerin hayatta kalma oranı -80°C'deki saklama koşullarında daha yüksek bulunmuştur. Ayrıca, lenfositlerin -80°C'de kriyoprezervasyon çözeltisi ile (%10 DMSO) saklanmasının lenfosit bütünlüğünü korumak için daha uygun bir strateji sağladığı söylenebilir.
Keywords
References
- 1. Ho, C.K., Choi, S.W., Siu, P.M., Benzie, I.F. (2011). Cryopreservation and storage effects on cell numbers and DNA damage in human lymphocytes. Biopreservation and Biobanking, 9(4), 343-347. [CrossRef]
- 2. Germann, A., Oh, Y.J., Schmidt, T., Schön, U., Zimmermann, H., von Briesen, H. (2013). Temperature fluctuations during deep temperature cryopreservation reduce PBMC recovery, viability and T-cell function. Cryobiology, 67(2), 193-200. [CrossRef]
- 3. Belloni, P., Meschini, R., Palitti, F. (2008). Effects of storage conditions of human whole blood on the viability of lymphocytes. International Journal of Radiation Biology, 84(7), 613-619. [CrossRef]
- 4. Allione, A., Porcedda, P., Russo, A., Ricceri, F., Simonelli, V., Minoprio, A., Guarrera, S., Pardini, B., Mazzei, F., Dogliotti, E., Giachino, C., Matullo, G. (2013). Effect of blood storage conditions on DNA repair capacity measurements in peripheral blood mononuclear cells. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, 749(1-2), 73-79. [CrossRef]
- 5. Jalla, S., Sazawal, S., Deb, S., Black, R. E., Das, S.N., Sarkar, A., Bhan, M.K. (2004). Enumeration of lymphocyte subsets using flow cytometry: Effect of storage before and after staining in a developing country setting. Indian Journal of Clinical Biochemistry, 19, 95-99. [CrossRef]
- 6. Duthie, S.J., Pirie, L., Jenkinson, A.M., Narayanan, S. (2002). Cryopreserved versus freshly isolated lymphocytes in human biomonitoring: Endogenous and induced DNA damage, antioxidant status and repair capability. Mutagenesis, 17(3), 211-214. [CrossRef]
- 7. Belloni, P., Pepe, G., Palitti, F. (2010). Effect of storage conditions of blood on radiation-induced chromosomal aberrations and apoptosis in human lymphocytes. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 695(1-2), 35-39. [CrossRef]
- 8. Park, M.S., Terasaki, P.I. (1974). Storage of human lymphocytes at room temperature. Transplantation, 18(6), 520-524. [CrossRef]
- 9. Stopford, C.R., Macqueen, J.M., Amos, D.B., Ward, F.E. (1972). Some variations in lymphocyte freezing methods which do not affect cell viability. Tissue Antigens, 2(1), 20-26. [CrossRef]
- 10. Vaught, J.B. (2006). Blood collection, shipment, processing, and storage. Cancer Epidemiology Biomarkers & Prevention, 15(9), 1582-1584. [CrossRef]
- 11. Ladeira, C., Koppen, G., Scavone, F., Giovannelli, L. (2019). The comet assay for human biomonitoring: effect of cryopreservation on DNA damage in different blood cell preparations. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 843, 11-17. [CrossRef]
- 12. Ekong, T., Kupek, E., Hill, A., Clark, C., Davies, A., Pinching, A. (1993). Technical influences on immunophenotyping by flow cytometry the effect of time and temperature of storage on the viability of lymphocyte subsets. Journal of Immunological Methods, 164(2), 263-273. [CrossRef]
There are 12 citations in total.
Details
| Primary Language | English |
|---|---|
| Subjects | Pharmaceutical Toxicology |
| Journal Section | Research Article |
| Authors | |
| Early Pub Date | May 4, 2025 |
| Publication Date | |
| Submission Date | December 7, 2024 |
| Acceptance Date | March 14, 2025 |
| Published in Issue | Year 2025 Volume: 49 Issue: 2 |