BibTex RIS Cite

The Effect of Different Cell Culture Surfaces on the Growth Proliferation Rate and Morphology of the Hepatocellular Carcinoma Cells

Year 2016, Volume: 8 Issue: 3, 12 - 17, 01.12.2016

Abstract

Aim: Cell culture surfaces effects the features of the cells. There are recent studies investigating the most suitable culture surfaces for the certain cell types. This study aims to examine the morphological and proliferative effects of the most three common cell culture surfaces, Schott glass, polystyrene, pyrex borosilicate glass, on the liver cancer cells. Materials And Methods: There were two experimental groups: For the first group; the sterilized D263M Schott glass coverslips were placed in pyrex borosilicate glass culture dishes. The second group contained polystyrene culture dishes which had sterilized D263M Schott glass coverslips like the first group. HepG2 cells, a hepatocellular carcinoma cell line, were cultured in those dishes. The images were daily captured under PrimoVert Invert phase contrast microscope. The areas containing cells were compared statistically. Results: No significant difference were observed between the cells on these three different culture surfaces. The statistically significant difference between the proliferation rate of the experimental groups were clear. Conclusions: Hepatocellular carcinoma cells prefers to adhere to the Schott glass, when it is compared with the other two surfaces. Iyt is likely that pyrex-borocilicate glass surfaces effects the proliferation rate. The chemical features of the cell culture surfaces are crucial for the succes of the experiments. Recent studies on the development of the most suitable cell culture environment for different cell types, like collagen application or matrigel usage, are extremely important for in vitro studies.

References

  • Santerre J.P, Woodhouse K, Laroche G, Labow R.S. Un- derstanding the biodegradation of polyurethanes: from classical implants to tissue engineering materials. Bioma- terials. 2005; 26: 7457-7470.
  • Bokhari M, Carnachan RJ, Cameron NR, Przyborski SA. Culture of HepG2 liver cells on three dimensional pol- ystyrene scaffolds enhances cell structure and function during toxicological challenge. J Anat. 2007; 211(4): 567-76.
  • Harrison RG. On the Stereotropism of Embryonic Cells. Science. 1911; 34(870): 279-81.
  • Berthier E, Young EW, Beebe D. Engineers are from PDMS-land, Biologists are from Polystyrenia. Lab on a chip. 2012; 12(7): 1224-37.
  • Amstein C.F, Hartman P.A. Adaption of Plastic Surfaces for Tissue Culture by Glow Discharge. J. Clinical Microbi- ology. 1975; 2: 46-54.
  • Curtis A.S.G, Forrester J.V, McInnes C, Lawrie F. Adhe- sion of Cells to Polystyrene Surfaces. J. Cell Biology. 1983; 97: 1500-1506.
  • Hudis, M. Plasma Treatment of Solid Materials. In: Hol- lahan, J.R.; Bell, A.T.; Ed. Techniques and Applications of Plasma Chemistry. John Wiley and Sons, New York. 1974: 113-147.
  • Ramsey W.S, Hertl W, Nowlan E.D, Binkowski N.J. Sur- face Treatments and Cell Attachment. In Vitro, Vol. 1984; 20: 802-808.
  • Bonfield TL, Colton E, Anderson JM. Plasma protein adsorbed biomedical polymers: activation of human monocytes and induction of interleukin 1. Journal of bi- omedical materials research. 1989; 23(6): 535-48.
  • Yun JK, DeFife K, Colton E, Stack S, Azeez A, Cahalan L, et al. Human monocyte/macrophage adhesion and cytokine production on surface-modified poly(tetrafluo- roethylene/hexafluoropropylene) polymers with and without protein preadsorption. Journal of biomedical materials research. 1995; 29(2): 257-68.
  • Reid LM, Rojkind M. New techniques for culturing differ- entiated cells: reconstituted basement membrane rafts. Methods in enzymology. 1979; 58:263-78.
  • Ryan J.A. Evolution of Cell Culture Surfaces. BioFiles. 2008; 3:8-21.
  • Matheson LA, McBane JE, Malowany JI, Santerre JP, Labow RS. Is cell culture stressful? Effects of degradable and nondegradable culture surfaces on U937 cell func- tion. BioTechniques. 2007; 42(6): 744, 6-50.
  • McNally A.K, Anderson J.M. Interleukin-4 induces foreign body giant cells from human monocytes/macrophages. Differential lymphokine regulation of macro- phage fu- sion leads to morphological variants of multinucleated giant cells. Am. J. Pathol. 1995; 147: 1487-1499.
  • Reid L.M, Rojkind M. New Techniques for Culturing Differentiated Cells: Reconstituted Basement Membrane Rafts. In Jakoby, W. B.; Pastan, I. H.; Ed. Cell Culture. Vol- ume 58 Methods in Enzymology, Chapter 21, Academic Press, New York. 1979; 263-278.
  • Murray LM, Nock V, Evans JJ, Alkaisi MM. The use of substrate materials and topography to modify growth patterns and rates of differentiation of muscle cells. Journal of biomedical materials research Part A. 2016; 104(7): 1638-45.

Karaciğer Kanser Hücrelerinin Büyüme, Çoğalma ve Morfolojileri Üzerine Farklı Kültür Yüzeylerinin Etkisi

Year 2016, Volume: 8 Issue: 3, 12 - 17, 01.12.2016

Abstract

Amaç: Hücrelerin farklı kültür yüzey koşullarında gelişim ve proliferasyon süreci değişkenlik göstermektedir. Son yıllarda yapılan çalışmalarla; hücre tipine özel en uygun kültür yüzeyleri saptanmaya çalışılmaktadır. Bu çalışmada; in vitro kültür deneylerinde en sık kullanılan Schott cam, polistiren, pyrex borosilikat cam yüzeylerin karaciğer kanser hücrelerinin çoğalmaları ve morfolojileri üzerine etkilerinin karşılaştırmalı incelenmesi amaçlanmıştır. Gereç ve Yöntem: İki farklı deney grubu oluşturuldu. Birinci grup için cam otoklavize edilebilen, pyrex borosilikat petri kapları sterilize edildi, sterilize edilmiş D263M Schott cam lameller yerleştirildi. İkinci deney grubu için ise, polistirenden yapılan steril kültür kapları kullanıldı. Birinci gruptakine benzer şekilde Schott cam lameller yerleştirildi. Hepatosellüler karsinoma tanısı konulmuş bir hastadan izole edilen altıncı pasaj HepG2 hücreleri ekildi. Hücrelerin günlük takipleri yapılarak PrimoVert Invert mikroskop ile farklı büyütmelerde canlı görüntüleri elde edildi. Çekilen fotoğraflar üzerinde hücrelerin bulunduğu alanların ölçümü yapılarak karşılaştırmalı değerlendirildi. Bulgular: Morfolojik açıdan Pyrex borosilikat cam, D263M Schott cam ve polistiren yüzeylerde büyüyen hücreler arasında belirgin bir farklılık gözlemlenmedi. Ancak hücrelerin kapladığı alanlar ölçülerek karşılaştırıldığında; sırasıyla D263M Schott cam, polistiren ve pyrex borosillicate cam kültür yüzeyler üzerinde çoğalma oranının istatistiksel olarak anlamlı şekilde azalmış olduğu görüldü. Sonuçlar: Hücreler, en çok D263M Schott cam üzerinde çoğalmayı tercih etmişlerdir. Hücre çoğalması en az pyrex-borosilikat cam yüzeyler üzerinde olmuştur. Sonuçlarımız; hücre kültür yüzeylerinin kimyasal özelliklerinin in vitro çalışmaların başarısını önemli şekilde etkileyebileceğini göstermektedir. Pyrex-borosilikat yüzeyde çoğalmadaki düşüklük; son yıllarda cam yüzeylerin hücre karakterine uygun kollojen, matrijel ve benzeri maddelerle kaplanması üzerine yapılan çalışmaların önemini ortaya çıkarmaktadır.

References

  • Santerre J.P, Woodhouse K, Laroche G, Labow R.S. Un- derstanding the biodegradation of polyurethanes: from classical implants to tissue engineering materials. Bioma- terials. 2005; 26: 7457-7470.
  • Bokhari M, Carnachan RJ, Cameron NR, Przyborski SA. Culture of HepG2 liver cells on three dimensional pol- ystyrene scaffolds enhances cell structure and function during toxicological challenge. J Anat. 2007; 211(4): 567-76.
  • Harrison RG. On the Stereotropism of Embryonic Cells. Science. 1911; 34(870): 279-81.
  • Berthier E, Young EW, Beebe D. Engineers are from PDMS-land, Biologists are from Polystyrenia. Lab on a chip. 2012; 12(7): 1224-37.
  • Amstein C.F, Hartman P.A. Adaption of Plastic Surfaces for Tissue Culture by Glow Discharge. J. Clinical Microbi- ology. 1975; 2: 46-54.
  • Curtis A.S.G, Forrester J.V, McInnes C, Lawrie F. Adhe- sion of Cells to Polystyrene Surfaces. J. Cell Biology. 1983; 97: 1500-1506.
  • Hudis, M. Plasma Treatment of Solid Materials. In: Hol- lahan, J.R.; Bell, A.T.; Ed. Techniques and Applications of Plasma Chemistry. John Wiley and Sons, New York. 1974: 113-147.
  • Ramsey W.S, Hertl W, Nowlan E.D, Binkowski N.J. Sur- face Treatments and Cell Attachment. In Vitro, Vol. 1984; 20: 802-808.
  • Bonfield TL, Colton E, Anderson JM. Plasma protein adsorbed biomedical polymers: activation of human monocytes and induction of interleukin 1. Journal of bi- omedical materials research. 1989; 23(6): 535-48.
  • Yun JK, DeFife K, Colton E, Stack S, Azeez A, Cahalan L, et al. Human monocyte/macrophage adhesion and cytokine production on surface-modified poly(tetrafluo- roethylene/hexafluoropropylene) polymers with and without protein preadsorption. Journal of biomedical materials research. 1995; 29(2): 257-68.
  • Reid LM, Rojkind M. New techniques for culturing differ- entiated cells: reconstituted basement membrane rafts. Methods in enzymology. 1979; 58:263-78.
  • Ryan J.A. Evolution of Cell Culture Surfaces. BioFiles. 2008; 3:8-21.
  • Matheson LA, McBane JE, Malowany JI, Santerre JP, Labow RS. Is cell culture stressful? Effects of degradable and nondegradable culture surfaces on U937 cell func- tion. BioTechniques. 2007; 42(6): 744, 6-50.
  • McNally A.K, Anderson J.M. Interleukin-4 induces foreign body giant cells from human monocytes/macrophages. Differential lymphokine regulation of macro- phage fu- sion leads to morphological variants of multinucleated giant cells. Am. J. Pathol. 1995; 147: 1487-1499.
  • Reid L.M, Rojkind M. New Techniques for Culturing Differentiated Cells: Reconstituted Basement Membrane Rafts. In Jakoby, W. B.; Pastan, I. H.; Ed. Cell Culture. Vol- ume 58 Methods in Enzymology, Chapter 21, Academic Press, New York. 1979; 263-278.
  • Murray LM, Nock V, Evans JJ, Alkaisi MM. The use of substrate materials and topography to modify growth patterns and rates of differentiation of muscle cells. Journal of biomedical materials research Part A. 2016; 104(7): 1638-45.
There are 16 citations in total.

Details

Primary Language Turkish
Journal Section Research Article
Authors

Hatice İsan This is me

Önder Tombuş This is me

Meltem Elif Göklü This is me

Kübra Tüfekçi This is me

Ranan Gülhan Aktaş This is me

Publication Date December 1, 2016
Published in Issue Year 2016 Volume: 8 Issue: 3

Cite

Vancouver İsan H, Tombuş Ö, Göklü ME, Tüfekçi K, Aktaş RG. Karaciğer Kanser Hücrelerinin Büyüme, Çoğalma ve Morfolojileri Üzerine Farklı Kültür Yüzeylerinin Etkisi. Maltepe tıp derg. 2016;8(3):12-7.