Harika Eylül ESMER DURUEL, Nazlı Sevim ÇAĞAN, Serra IŞIK, Figen Esin KAYHAN

Hücre Kültürlerine Genel Bakış

Hücre kültürü yöntemi, biyoteknolojik bağlamda geliştirilen bir yöntem olup hücrelerin laboratuvar şartlarında ve özel koşullar altında, doğal ortamlarının dışında yetiştirilmelerini ifade eder. İlk olarak yetişkin kurbağa sinir liflerinin, lenf sıvıları içerisinde büyütülmesi ile hayatımıza girmiştir. Geçmişten günümüze çalışılmakta olan hücre hatlarının gereksinimlerinin birbirlerinden farklı oldukları belirlenmiştir. Bu farklılıklar göz önünde bulundurularak doku, vasat ve ortam koşulları değiştirilmiş ve günümüzde kullanımı devam etmekte olan farklı hücre hatları ile kültür yöntemleri bulunmuştur. Günümüzde farmasötik, tıbbi ve biyolojik araştırmalarda çeşitli hücre hatları kullanılmaktadır. Bu hücre hatları, hücre biyolojisi ile doku morfolojisinin anlaşılması ve bu sayede doku mühendisliğinin geliştirilmesi için kullanılmaktadır. Genel olarak iki boyutlu hücre kültürü modeli kullanılıyor olsa da son zamanlarda üç boyutlu hücre kültürü modelleri de tercih edilmektedir. Besiyerleri farklı ihtiyaçlara yönelik hizmet etseler de genel olarak ortak bazı bileşenlerin hücre kültürü ortamları için son derece önemli olduğu bilinmektedir. Bu ortak bileşenler incelendiğinde özellikle karbonhidrat kaynakları ve serum ilk sıralarda yer almaktadır. Bunun en temel nedeni karbonhidratların temel enerji kaynaklarını oluşturması ve serumun ise gerekli büyüme faktörlerinin yanı sıra farklı hormon ve bileşenler içermesidir. Hücre kültürü uygulamalarında vasat kimyasal bileşenleri kadar hücrelerin bulunduğu ortamın çevresel faktörleri de hücre büyümesi ve devamlılığı açısından önemlidir. Ayrıca hücre kültürü yöntemlerinde hijyen konusuna hassasiyet gösterilmelidir. Çünkü kültür ortamı, kimyasal bileşenler ve çevresel faktörler birlikte ele alındığında aseptik koşullarda çalışılmaz ise kontaminasyon riski ortaya çıkmaktadır. Bu derlemede hücre kültürü yöntem ve uygulamalarına genel bir bakış açısı sunulacaktır.

Anahtar Kelimeler:

Hücre kültürü, hücre vasatı, kontaminasyon, hücre hattı, büyüme faktörleri

Cell culture method is a method developed in a biotechnological context and refers to the cultivation of cells outside of their natural environment under laboratory conditions and special conditions. It first came into our lives with the enlargement of adult frog nerve fibers in lymph fluids. Studies have shown that the needs of cell lines are different from each other. Considering these differences, tissue, media and environment conditions have been changed and different cell lines and culture methods that are in use today have been found. Various cell lines are currently used in pharmaceutical, medical and biological research. Cell culture studies are carried out to understand the biological structure and tissue morphology of the cell, thus improving tissue engineering. Although two-dimensional cell culture models are generally used, three-dimensional cell culture models are also preferred recently. Although the media serve for different needs, it has been found that some common components are extremely important for cell culture media in general. When these common components are examined, especially carbohydrate sources and serum are in the first place. Since carbohydrates are the main energy sources and serum contains different hormones and components as well as necessary growth factors, it is added to the medium. In cell culture practices, the environmental factors of the environment where the cells are located and the chemi-cal components of the environment are important for cell growth and continuity. In addition, cell culture methods should be sensitive to hygiene. Because when the culture environment, chemical components and environmental factors are taken together, if the aseptic conditions are not studied, there is a risk of contamination. In this review, an overview of cell culture methods and applications will be presented.

Keywords:

Cell culture, cell medium, contamination, cell line, growth factors,

PDF

___

Ackermann T and Tardito S (2019). Cell culture medium formulation and ıts ımplications in cancer metabolism. Trends in Cancer 5(6), 329–332. https://doi.org/10.1016/j.trecan.2019.05.004
Bryant KL, Stalnecker CA, Zeitouni D, Klomp JE, Peng S, Tikunov AP, Gunda V, Pierobon M, Waters AM, George SD, Tomar G, Papke B, Hobbs GA, Yan L, Hayes TK, Diehl JN, Goode GD, Chaika NV, Wang Y, Der CJ (2019). Combination of ERK and autophagy inhibition as a treatment approach for pancreatic cancer. Nature Medicine, 25(4), 628–640. https://doi.org/10.1038/s41591-019-0368-8
Carrel A (1912). On the permanent lıfe of tıssues outsıde of the organısm. The Journal of Experimental Medicine, 15(5), 516–528. https://doi.org/10.1084/jem.15.5.516
Carrel A and Burrows MT (1911). Cultıvatıon of tıssues ın vıtro and ıts technıque. The Journal of Experimental Medicine, 13(3), 387–396. https://doi.org/10.1084/jem.13.3.387
Chu GL and Dewey WC (1987). Effect of hyperthermia on intracellular pH in Chinese hamster ovary cells. Radiation Research, 110(3), 439–449.
Chung C and Burdick JA (2008). Engineering cartilage tissue. Advanced Drug Delivery Reviews, 60(2), 243–262. https://doi.org/10.1016/j.addr.2007.08.027
Combs GF (2012). How to Use This Book. Academic Press. https://doi.org/https://doi.org/10.1016/B978-0-12-381980-2.00032-3
Donaldson CD and Bishop KN (2015). Cell culture. British Journal of Hospital Medicine, 76(1), C2–C5. https://doi.org/10.12968/hmed.2015.76.1.C2
Dulbecco R and Freeman G (1959). Plaque production by the polyoma virus. Virology, 8(3), 396–397. https://doi.org/10.1016/0042-6822(59)90043-1
Eagle H (1955). Nutrition needs of mammalian cells in tissue culture. Science (New York, N.Y.), 122(3168), 501–514. https://doi.org/10.1126/science.122.3168.501
Earle WR, Schilling EL, Stark TH, Straus NP, Brown MF, Shelton E (1943). IV. The mouse fibroblast cultures and changes seen in the living cells. Journal of the National Cancer Institute, 4(2), 165–212.
Fischer A (1948). Amino-acid metabolism of tissue cells in vitro. Nature, 161(4104), 1008. https://doi.org/10.1038/1611008a0
Gey GO (1952). Tissue culture studies of the proliferative capacity of cervical carcinoma and normal epithelium. Cancer Res., 12, 264–265.
Gola J, Skubis A, Sikora B, Krusznıewska-Rajs C, Adamska J, Mazurek U, Strzałka-Mrozık B, Czernel G, Gagoś M (2015). Expression profiles of genes related to melatonin and oxidative stress in human renal proximal tubule cells treated with antibiotic amphotericin B and its modified forms. Turkısh Journal Of Bıology, 39, 856–864. https://doi.org/10.3906/biy-1505-52
Ham RG (1963). An improved nutrient solution for diploid Chinese hamster and human cell lines. Experimental Cell Research, 29(3), 515–526. https://doi.org/10.1016/S0014-4827(63)80014-2
Harrison RG (1907). Observations on the living developing nerve fiber. Proceedings of the Society for Experimental Biology and Medicine, 4(1), 140–143. https://doi.org/10.3181/00379727-4-98
Hassan SN and Ahmad F (2020). The relevance of antibiotic supplements in mammalian cell cultures: Towards a paradigm shift. Gulhane Medical Journal, 62(4), 224–230. https://doi.org/10.4274/gulhane.galenos.2020.871
Howorth PJ (1975). The physiological assessment of acid-base balance. British Journal of Diseases of the Chest, 69(2), 75–102.
Jedrzejczak-Silicka M (2017). History of cell culture. New Insights into Cell Culture Technology, 1–42. https://doi.org/10.5772/66905
Kapałczyńska M, Kolenda T, Przybyła W, Zajączkowska M, Teresiak A, Filas V, Ibbs M, Bliźniak R, Łuczewski Ł, Lamperska K (2018). 2D and 3D cell cultures – a comparison of different types of cancer cell cultures. Archives of Medical Science, 14(4), 910–919. https://doi.org/10.5114/aoms.2016.63743
Krattenmacher F, Heermann T, Calvet A, Krawczyk B, Noll T (2018). Effect of manufacturing temperature and storage duration on stability of chemically defined media measured with LC-MS/MS: Impact of manufacturing and storage on CDM components measured with LC-MS/MS. Journal of Chemical Technology ve Biotechnology, 94. https://doi.org/10.1002/jctb.5861
Lewis MR and Lewis W (1911). The cultıvatıon of tıssues from chıck embryos ın solutıons of NaCI, CaC12, KCl AND NaHC03. The Anatomical Record, 5(6), 277–293. https://doi.org/10.1126/science.39.994.107
Mather J and Sato G (1977). Hormones and growth factors in cell cultures: problems and perspectives. Cell, Tissue, and Organ Cultures in Neurobiology (ss. 619–630). Elsevier.
McGillicuddy N, Floris P, Albrecht S, Bones J (2018). Examining the sources of variability in cell culture media used for biopharmaceutical production. Biotechnology Letters, 40(1), 5–21. https://doi.org/10.1007/s10529-017-2437-8
Michl J, Park KC, Swietach P (2019). Evidence-based guidelines for controlling pH in mammalian live-cell culture systems. Communications Biology, 2(1), 1–12. https://doi.org/10.1038/s42003-019-0393-7
Nims RW and Harbell JW (2017). Best practices for the use and evaluation of animal serum as a component of cell culture medium. In Vitro Cellular and Developmental Biology - Animal, 53(8), 682–690. https://doi.org/10.1007/s11626-017-0184-8
Price P(2017). Best practices for media selection for mammalian cells. In Vitro Cellular and Developmental Biology - Animal, 53(8), 673–681. https://doi.org/10.1007/s11626-017-0186-6
Ringer S (1882). Concerning the ınfluence exerted by each of the constituents of the blood on the contraction of the ventricle. The Journal of Physiology, 3(5–6), 380–393. https://doi.org/10.1113/jphysiol.1882.sp000111
Rothblat G (2012). Growth, Nutrition, and Metabolism of Cells In Culture V3 (C. 3). Elsevier.
Ryu AH, Eckalbar WL, Kreimer A, Yosef N, Ahituv N (2017). Use antibiotics in cell culture with caution: Genome-wide identification of antibiotic-induced changes in gene expression and regulation. Scientific Reports, 7(1), 1–9. https://doi.org/10.1038/s41598-017-07757-w
Salazar A, Keusgen M, Von Hagen J (2016). Amino acids in the cultivation of mammalian cells. Amino Acids, 48(5), 1161–1171. https://doi.org/10.1007/s00726-016-2181-8
Schwartz PB and Ronnekleiv-Kelly SM (2019). Effective Cell Culture. 157–169. https://doi.org/10.1007/978-3-030-14644-3_10
Shipman CJ (1969). Evaluation of 4-(2-hydroxyethyl)-1-piperazineëthanesulfonic acid (HEPES) as a tissue culture buffer. Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.), 130(1), 305–310. https://doi.org/10.3181/00379727-130-33543
Sung YH, Lim SW, Chung JY, Lee GM (2004). Yeast hydrolysate as a low-cost additive to serum-free medium for the production of human thrombopoietin in suspension cultures of Chinese hamster ovary cells. Applied Microbiology and Biotechnology, 63(5), 527–536. https://doi.org/10.1007/s00253-003-1389-1
van der Valk J, Brunner D, De Smet K, Fex Svenningsen A, Honegger P, Knudsen LE, Lindl T, Noraberg J, Price A, Scarino ML, Gstraunthaler G (2010). Optimization of chemically defined cell culture media--replacing fetal bovine serum in mammalian in vitro methods. Toxicology in Vitro : An International Journal Published in Association with BIBRA, 24(4), 1053–1063. https://doi.org/10.1016/j.tiv.2010.03.016
Vogelaar JPM and Erlichman E (1933). A feeding solution for cultures of human fibroblasts. Am J Cancer, 18, 28–38.
Wahrheit J, Nicolae A, Heinzle E (2014). Dynamics of growth and metabolism controlled by glutamine availability in Chinese hamster ovary cells. Applied Microbiology and Biotechnology, 98(4), 1771–1783. https://doi.org/10.1007/s00253-013-5452-2
Yao T, and Asayama Y (2017). Animal-cell culture media: History, characteristics, and current issues. Reproductive Medicine and Biology, 16(2), 99–117. https://doi.org/10.1002/rmb2.12024