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293T İnsan Renal Hücrre Dizisinde LPS Konsantrasyonlarının Yeniden Değerlendirilmesi

Year 2016, , 20 - 22, 31.01.2016

Aris Çakiris Atilla Cakar Neslihan Abacı Sema Ekmekci Zeliha Emrence , Duran Üstek

https://doi.org/10.30934/kusbed.358484

Abstract

Amaç: Hücrelerin LPS (Lipopolisakkarit)’ye enflamatuvaryanıtları in-vitro deneylerde sıklıkla kullanılmaktadır. Buna rağmen hücre kültürü çalışmaları için LPS ’nin doğru dozunun belirlenmesinde zorluklarla karşılaşılmaktadır. Maksimum yanıt veren LPS konsantrasyonunun belirlenmesi in vitro enflamatuvar deneyler için kritik bir noktadır. Bu çalışmanın amacı LPS’nin 293T insan renal hücre dizisindeki konsantrasyon bağımlı etkisinin yeniden değerlendirilmesidir.

Yöntem: Bu çalışmada LPS ile stimule edilmiş 293T hücre hattı Xcelligence Real-Time Cell Analyzer (RTCA) sisteminde incelendi. LPS ’nin artmış konsantrasyonlarının 293T hücre hattı üzerindeki etkisi RTCA cihazında hücre proliferasyonu ölçümleri yapılarak araştırıldı.

Bulgular: Sonuçlarımıza göre 2, 4 ve 8 μg’lık LPS konsantrasyonlarını hücre bölünmesini inhibe ederek hücreleri kararlı duruma yönlendirdiğini gösterdi. 1 μg ’lık LPS konsantrasyonu hücreleri mitotik faza sürüklerken daha düşük LPS konsantrasyonlarının ise hücre proliferasyonu üzerinde bir etki göstermedi.

Sonuç: Bu çalışmadaki sonuçlara göre LPS konsantrasyonlarının hücre proliferasyonu üzerinde farklı etkilerinin olduğu ve deneysel çalışmalardan önce LPS konsantrasyonlarının optimize edilmesi gerektiği gösterilmiştir.

Keywords

lipopolisakkarit LPS hücre proliferasyonu enflamasyon

References

  • Ruiz N, Kahne D, Silhavy T.J. Transport of lipopolysaccharide across the cellenvelope: the long road of discovery. Nat Rev Microbiol. 2009; 7(9): 677–683.
  • Fumarola D., Jirillo E J. Contamination of con Apreperations. Immunol. 1976.116:1197.
  • Fumarola D., Jirillo E J.Endotoxin contamination of some commercial preparationused in experimental research. Biomedical applications 1979.29:379-385.
  • Fumarola,D.,E. Jirillo,G.Miragliotta,and E. Magliulo. Influence oflipopolysaccharide from.Pasteurellamultocida on fibroblasts cultured in vitro. IRCSMed Sci. 1982; 10: 647-648
  • Del Liano, A.M. and.Lavergne, J.A.Submitogenic doses of lipopolysaccharide alter the patterns of nuclear granularity and cell proliferation in human mixed lymphocyte cultures. Transplantation Proceedings. 1991. 23:1766-1770.
  • Abassi YA, Xi B, Zhang W, et al. Kinetic cell-based morphological screening:prediction of mechanism of compound action and off-target effects. ChemBiol2009;16(7):712-23.
  • Kulahava TA, Semenkova GN, Kvacheva ZB, et al. Effects of peroxynitrite andlipopolysaccharide on mitotic activity of C6 glioma cells.NeurosciLett. 2006;398(3):286-90.
  • Moore RN, Steeg PS, Männel DN, Mergenhagen SE. Role of lipopolysaccharide inregulating colony-stimulating factor-dependent macrophage proliferation in vitro.Infect Immun. 1980 ;30(3):797-804.

Re-evaluation Of LPS Concentrations On The 293T Human Renal Cell Line

Year 2016, , 20 - 22, 31.01.2016

Aris Çakiris Atilla Cakar Neslihan Abacı Sema Ekmekci Zeliha Emrence , Duran Üstek

https://doi.org/10.30934/kusbed.358484

Abstract

Objective: The inflammatory responsiveness of the cells to Lipopolysaccharides (LPS) is commonly used for in vitro experiments. However, the correct dose of the LPS for cell line experiments is elusive. The LPS concentration that gives the maximal response is a critical point of in vitro inflammatory experiments. The aim of this study was to reevaluate the concentration dependent effect of LPS on the 293T human renal cell line.

Methods: We evaluated the cell-detachment assay of LPS-stimulated 293T cell line monitored by xCELLigence Real-Time Cell Analyzer (RTCA) system. We applied increasing concentration of LPS followed by Roche xCELLigence Instrument based on the Real-Time Cell Analysis System.

Results: Our results demonstrated that the 2, 4 and 8μg of LPS inhibit cell division which diverts cells to a steady-state phase, 1 μg acts as mitogen. Lower concentrations are no effect on cells.

Conclusion: These work showed that LPS concentrations had various effects on cell proliferation and need to be estimated for each experiment before carry out the experiments.

Keywords

Lipopolysaccharides LPS cell proliferation inflammation

References

  • Ruiz N, Kahne D, Silhavy T.J. Transport of lipopolysaccharide across the cellenvelope: the long road of discovery. Nat Rev Microbiol. 2009; 7(9): 677–683.
  • Fumarola D., Jirillo E J. Contamination of con Apreperations. Immunol. 1976.116:1197.
  • Fumarola D., Jirillo E J.Endotoxin contamination of some commercial preparationused in experimental research. Biomedical applications 1979.29:379-385.
  • Fumarola,D.,E. Jirillo,G.Miragliotta,and E. Magliulo. Influence oflipopolysaccharide from.Pasteurellamultocida on fibroblasts cultured in vitro. IRCSMed Sci. 1982; 10: 647-648
  • Del Liano, A.M. and.Lavergne, J.A.Submitogenic doses of lipopolysaccharide alter the patterns of nuclear granularity and cell proliferation in human mixed lymphocyte cultures. Transplantation Proceedings. 1991. 23:1766-1770.
  • Abassi YA, Xi B, Zhang W, et al. Kinetic cell-based morphological screening:prediction of mechanism of compound action and off-target effects. ChemBiol2009;16(7):712-23.
  • Kulahava TA, Semenkova GN, Kvacheva ZB, et al. Effects of peroxynitrite andlipopolysaccharide on mitotic activity of C6 glioma cells.NeurosciLett. 2006;398(3):286-90.
  • Moore RN, Steeg PS, Männel DN, Mergenhagen SE. Role of lipopolysaccharide inregulating colony-stimulating factor-dependent macrophage proliferation in vitro.Infect Immun. 1980 ;30(3):797-804.
There are 8 citations in total.

Details

Subjects Health Care Administration
Journal Section Original Article
Authors

Aris Çakiris This is me

Atilla Cakar This is me

Neslihan Abacı This is me

Sema Ekmekci This is me

Zeliha Emrence

Duran Üstek This is me

Publication Date January 31, 2016
Submission Date November 18, 2015
Acceptance Date December 20, 2015
Published in Issue Year 2016

Cite

APA Çakiris, A., Cakar, A., Abacı, N., Ekmekci, S., et al. (2016). Re-evaluation Of LPS Concentrations On The 293T Human Renal Cell Line. Kocaeli Üniversitesi Sağlık Bilimleri Dergisi, 2(1), 20-22. https://doi.org/10.30934/kusbed.358484
AMA Çakiris A, Cakar A, Abacı N, Ekmekci S, Emrence Z, Üstek D. Re-evaluation Of LPS Concentrations On The 293T Human Renal Cell Line. KOU Sag Bil Derg. January 2016;2(1):20-22. doi:10.30934/kusbed.358484
Chicago Çakiris, Aris, Atilla Cakar, Neslihan Abacı, Sema Ekmekci, Zeliha Emrence, and Duran Üstek. “Re-Evaluation Of LPS Concentrations On The 293T Human Renal Cell Line”. Kocaeli Üniversitesi Sağlık Bilimleri Dergisi 2, no. 1 (January 2016): 20-22. https://doi.org/10.30934/kusbed.358484.
EndNote Çakiris A, Cakar A, Abacı N, Ekmekci S, Emrence Z, Üstek D (January 1, 2016) Re-evaluation Of LPS Concentrations On The 293T Human Renal Cell Line. Kocaeli Üniversitesi Sağlık Bilimleri Dergisi 2 1 20–22.
IEEE A. Çakiris, A. Cakar, N. Abacı, S. Ekmekci, Z. Emrence, and D. Üstek, “Re-evaluation Of LPS Concentrations On The 293T Human Renal Cell Line”, KOU Sag Bil Derg, vol. 2, no. 1, pp. 20–22, 2016, doi: 10.30934/kusbed.358484.
ISNAD Çakiris, Aris et al. “Re-Evaluation Of LPS Concentrations On The 293T Human Renal Cell Line”. Kocaeli Üniversitesi Sağlık Bilimleri Dergisi 2/1 (January 2016), 20-22. https://doi.org/10.30934/kusbed.358484.
JAMA Çakiris A, Cakar A, Abacı N, Ekmekci S, Emrence Z, Üstek D. Re-evaluation Of LPS Concentrations On The 293T Human Renal Cell Line. KOU Sag Bil Derg. 2016;2:20–22.
MLA Çakiris, Aris et al. “Re-Evaluation Of LPS Concentrations On The 293T Human Renal Cell Line”. Kocaeli Üniversitesi Sağlık Bilimleri Dergisi, vol. 2, no. 1, 2016, pp. 20-22, doi:10.30934/kusbed.358484.
Vancouver Çakiris A, Cakar A, Abacı N, Ekmekci S, Emrence Z, Üstek D. Re-evaluation Of LPS Concentrations On The 293T Human Renal Cell Line. KOU Sag Bil Derg. 2016;2(1):20-2.