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Farklı Hücre Hatlarında Besiyeri ve FBS’in Hücre Proliferasyonu Üzerindeki Etkilerinin İncelenmesi

Yıl 2022, Cilt: 17 Sayı: 1, 55 - 64, 27.05.2022
https://doi.org/10.29233/sdufeffd.927126

Öz

Hücre kültürü, laboratuvar şartlarında hücrelerin canlılığının ve proliferasyonunun devam ettirilmesi için oluşturulan yapay, steril bir ortamdır. Bu yapay ortam şartlarında, hücrelerin büyümelerinin desteklenmesi amacıyla besiyerine ek olarak fetal bovine serum (FBS) gibi hücre proliferasyonunu arttırıcı maddeler de eklenmektedir. Hücreler uygun besiyeri ve FBS oranı kullanıldığı takdirde daha etkili bir şekilde prolifere olabilmektedir. Bu çalışmada, farklı besiyeri çeşitlerinin (Dulbecco's Modified Eagle's Medium (DMEM), RPMI-1640, DMEM High-Glucose ve DMEM/F12) ve farklı FBS oranlarının (%0, %5, %10, %15, %20, %25) belirlenen hücre hatlarındaki (L929, J774, Arpe-19, RVECs, AU-565, MCF-7, HEK293T) hücre proliferasyonuna etkileri incelendi. Hücre canlılık analizleri MTT testi ile yapıldı. MTT testi sonuçlarına göre belirlenen hücre hatlarının prolifere olacağı en uygun besiyeri çeşidi ve kullanılacak FBS oranları belirlendi. Bu çalışmayla birlikte, belirtilen hücre hatlarının birçok besiyeri çeşidinde ve farklı FBS oranlarında prolifere olabileceği gözlemlenmiştir. Elde edilen veriler dikkate alındığında, farklı hücre hatlarında optimum hücre proliferasyonun gerçekleştiği besiyeri çeşidi ve maliyeti yüksek ürünlerden biri olan FBS’in optimum hücre proliferasyonunu sağladığı ideal düzeydeki kullanım oranları saptanmıştır.

Kaynakça

  • [1] D.E. Lynn, Encylopedia of insects. Academic Press, 2009, ch 39.
  • [2] C. Eskes, A.C. Boström, G. Bowe, S. Coecke, T. Hartung, G. Hendriks D. Pamies, A. Piton, and C. Rovida, ‘‘Good cell culture pratices & in vitro toxicology,’’ Toxicology in Vitro, 45, 272-277, 2017.
  • [3] D.F. Gilbert and M. Boutros, ‘‘A protocol for a high-throughput multiplex cell viability assay,’’ Methods Mol Biol., 1470, 75-84, 2016.
  • [4] T. Ackermann, and S. Tardito, ‘‘Cell culture medium formulation and its implications in cancer metabolism,’’ Trends in Cancer, 5(6), 329-332, 2019.
  • [5] C.P. Segeritz, and L. Vallier, Basic Science Methods for Clinical Researchers. Academic Press, 2017, ch 9.
  • [6] M.A. Schwartz, G. Both, and C. Lechene, ‘‘Effect of cell spreading oc cytoplasmic pH in normal and transformed fibroblasts,’’ Proc Natl Sci U S A, 86(12), 4525-4529, 1989.
  • [7] C.E. Jochems, J.B. Valk, F.R. Stafleu, V. Baumans, ‘‘The use of fetal bonine serum: ethical or scientific problem,’’ Altern Lab Anim, 30(2), 219-227, 2002.
  • [8] T. Yao and Y. Asayama, “Animal-cell culture media: history, characteristics, and current issues,” Reprod. Med. Biol., 16(2), 99-117, 2017.
  • [9] T. Yao and Y. Asayama, “Human preimplantation embryo culture media: past, present, and future,” J. Mamm. Ova Res., 33(1), 17-34, 2016.
  • [10] G. Gstraunthaler, T. Lindl and J. Van Der Valk, “A plea to reduce or replace fetal bovine serum in cell culture media,” Cytotechnology, 65(5), 791-793, 2013.
  • [11] J.V. der Valk, K. Bieback, C. Buta, B. Cochrane, W.G. Dirks, J. Fu, J.J Hickman, C. Hohensee, R. Kolar, M. Liebsch, F. Pistollato, M. Schulz, D. Thieme, T. Weber, J. Wiest, S. Winkler and G. Gstraunthaler, “Fetal Bovine Serum (FBS): Past-Present-Future,” Altex, 35(1), 99-118, 2018.
  • [12] P. Pazos, M. Boveri, A. Gennari, J. Casado, F. Fernandez and P. Prieto, “Culturing cells without serum: lessons learnt using molecules of plant origin,” Altex, 21(2), 67-72, 2004.
  • [13] T.T. Cao and Y.Q. Zhang, “Viability and proliferation of L929, tumour and hybridoma cells in the culture media containing sericin protein as a supplement or serum substitute,” Appl. Microbiol. Biotechnol., 99(17) 7219-7228, 2015.
  • [14] Z. Wei, A.O. Batagov, D. R. Carter and A. M. Krichevsky, “Fetal Bovine Serum RNA interferes with the cell culture derived extracellular RNA,” Sci Rep. 6(1), 1-6, 2016.
  • [15] A. Wojdani, E.A. Stein, L.J. Alfred and E.L. Cooper, “Mitogenic effect of earthworm (Lumbricus terrestris) coelomic fluid on mouse and human lymphocytes,” Immunobiology, 166(2), 157-167, 1984.

Investigation of the Effects of Medium and FBS on Cell Proliferation in Different Cell Lines

Yıl 2022, Cilt: 17 Sayı: 1, 55 - 64, 27.05.2022
https://doi.org/10.29233/sdufeffd.927126

Öz

Cell culture is an artificial, sterile environment created to maintain the viability and proliferation of cells under laboratory conditions. In these artificial environment conditions, in addition to the medium, cell proliferation enhancing substances such as Fetal bovine serum (FBS) are added to support the growth of cells. Cells can proliferate more effectively if appropriate medium and FBS ratio are used. In this study, it was determined that different media types (Dulbecco's Modified Eagle's Medium (DMEM), RPMI-1640, DMEM High-Glucose and DMEM/F12) and different FBS ratios (0%, 5%, 10%, 15%, 20%, 25%) effects on cell proliferation in determined cell lines (L929, J774, Arpe-19, RVECs, AU-565, MCF-7, HEK293T) were examined. Cell viability analyzes were performed with the MTT test. According to the results of the MTT test, the most suitable medium type on which the cell lines will proliferate and the FBS ratios to be used were determined. With this study, it was determined that the specified cell lines could proliferate in many media types and at different FBS ratios. When cell studies are carried out considering the study data, the production of cells will be carried out using the most effective medium type for different cell lines and the FBS ratio at which optimum cell proliferation occurs. In this way, it is aimed to produce cells by using FBS, which is one of the most costly products in cell studies, at the most ideal level.

Kaynakça

  • [1] D.E. Lynn, Encylopedia of insects. Academic Press, 2009, ch 39.
  • [2] C. Eskes, A.C. Boström, G. Bowe, S. Coecke, T. Hartung, G. Hendriks D. Pamies, A. Piton, and C. Rovida, ‘‘Good cell culture pratices & in vitro toxicology,’’ Toxicology in Vitro, 45, 272-277, 2017.
  • [3] D.F. Gilbert and M. Boutros, ‘‘A protocol for a high-throughput multiplex cell viability assay,’’ Methods Mol Biol., 1470, 75-84, 2016.
  • [4] T. Ackermann, and S. Tardito, ‘‘Cell culture medium formulation and its implications in cancer metabolism,’’ Trends in Cancer, 5(6), 329-332, 2019.
  • [5] C.P. Segeritz, and L. Vallier, Basic Science Methods for Clinical Researchers. Academic Press, 2017, ch 9.
  • [6] M.A. Schwartz, G. Both, and C. Lechene, ‘‘Effect of cell spreading oc cytoplasmic pH in normal and transformed fibroblasts,’’ Proc Natl Sci U S A, 86(12), 4525-4529, 1989.
  • [7] C.E. Jochems, J.B. Valk, F.R. Stafleu, V. Baumans, ‘‘The use of fetal bonine serum: ethical or scientific problem,’’ Altern Lab Anim, 30(2), 219-227, 2002.
  • [8] T. Yao and Y. Asayama, “Animal-cell culture media: history, characteristics, and current issues,” Reprod. Med. Biol., 16(2), 99-117, 2017.
  • [9] T. Yao and Y. Asayama, “Human preimplantation embryo culture media: past, present, and future,” J. Mamm. Ova Res., 33(1), 17-34, 2016.
  • [10] G. Gstraunthaler, T. Lindl and J. Van Der Valk, “A plea to reduce or replace fetal bovine serum in cell culture media,” Cytotechnology, 65(5), 791-793, 2013.
  • [11] J.V. der Valk, K. Bieback, C. Buta, B. Cochrane, W.G. Dirks, J. Fu, J.J Hickman, C. Hohensee, R. Kolar, M. Liebsch, F. Pistollato, M. Schulz, D. Thieme, T. Weber, J. Wiest, S. Winkler and G. Gstraunthaler, “Fetal Bovine Serum (FBS): Past-Present-Future,” Altex, 35(1), 99-118, 2018.
  • [12] P. Pazos, M. Boveri, A. Gennari, J. Casado, F. Fernandez and P. Prieto, “Culturing cells without serum: lessons learnt using molecules of plant origin,” Altex, 21(2), 67-72, 2004.
  • [13] T.T. Cao and Y.Q. Zhang, “Viability and proliferation of L929, tumour and hybridoma cells in the culture media containing sericin protein as a supplement or serum substitute,” Appl. Microbiol. Biotechnol., 99(17) 7219-7228, 2015.
  • [14] Z. Wei, A.O. Batagov, D. R. Carter and A. M. Krichevsky, “Fetal Bovine Serum RNA interferes with the cell culture derived extracellular RNA,” Sci Rep. 6(1), 1-6, 2016.
  • [15] A. Wojdani, E.A. Stein, L.J. Alfred and E.L. Cooper, “Mitogenic effect of earthworm (Lumbricus terrestris) coelomic fluid on mouse and human lymphocytes,” Immunobiology, 166(2), 157-167, 1984.
Toplam 15 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Yapısal Biyoloji
Bölüm Makaleler
Yazarlar

Murat Ihlamur 0000-0002-0458-5638

Buşra Akgül 0000-0002-3566-8874

Emrah Şefik Abamor 0000-0002-9174-4528

Yayımlanma Tarihi 27 Mayıs 2022
Yayımlandığı Sayı Yıl 2022 Cilt: 17 Sayı: 1

Kaynak Göster

IEEE M. Ihlamur, B. Akgül, ve E. Ş. Abamor, “Farklı Hücre Hatlarında Besiyeri ve FBS’in Hücre Proliferasyonu Üzerindeki Etkilerinin İncelenmesi”, Süleyman Demirel University Faculty of Arts and Science Journal of Science, c. 17, sy. 1, ss. 55–64, 2022, doi: 10.29233/sdufeffd.927126.