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The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Characterization of HEK293 and Namalwa Cell Cultures by Using Design of Experiment

실험계획법을 이용한 HEK293 및 Namalwa 세포배양 특성 규명

  • Kang, Kyung-Ho (Department of Biological Engineering, Inha University) ;
  • Seo, Joon-Serk (Department of Biological Engineering, Inha University) ;
  • Kim, Dong-Il (Department of Biological Engineering, Inha University)
  • Received : 2012.06.14
  • Accepted : 2012.06.26
  • Published : 2012.06.30
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Abstract

Various human host cell lines, which are more effective than the other original human cell lines, have been developed and used. Highly efficient human cell line can be obtained from the fusion between human embryonic kidney 293 (HEK293) and human Burkitt's lymphoma cells (Namalwa). Fused cell line has the advantages of both cell lines such as the high transfection efficacy of HEK293 cells and the constitutive expression of Epstein-Barr virus (EBV) genome which is related with high expression of target protein and anti-apoptotic growth of Namalwa cells. In this study, characterization of two original cell lines was performed by using design of experiment (DOE) considering cell maintenance, media development, optimization of culture condition, and scale-up. The formation of aggregates was apparent with high glutamine concentration at more than 6 mM. Supplementation of hydrolysates showed positive effects on the growth performances of HEK293 cells. On the contrary, Namalwa cells showed negative results. It was confirmed that Namalwa cells were more sensitive to lower temperature at $35^{\circ}C$ and hyperosmotic condition over 260 mOsm/kg. In addition, both cell lines showed limited growth in 3-L bioreactor due to shear stress.

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References

  1. Butler, M. (2005) Animal cell cultures: recent achievements and perspectives in the production of biopharmaceuticals. Appl. Microbiol. Biotechnol. 68: 283-291. https://doi.org/10.1007/s00253-005-1980-8
  2. Cho, M. S., H. Yee, and S. Chan (2002) Establishment of a human somatic hybrid cell line for recombinant protein production. J. Biomed. Sci. 9: 631-638. https://doi.org/10.1007/BF02254991
  3. Cho, M. S., H. Yee, C. Brown, B. Mei, C. Mirenda, and S. Chan (2003) Versatile expression system for rapid and stable production of recombinant proteins. Biotechnol. Prog. 19: 229-232. https://doi.org/10.1021/bp0255964
  4. Mei, B., Y. Chen, J. Chen, C. Q. Pan, and J. E. Murphy (2006) Expression of human coagulation factor VIII in a human hybrid cell line, HKB11. Mol. Biotechnol. 34: 165-178. https://doi.org/10.1385/MB:34:2:165
  5. Klein, G., E. Klein, and E. Kashuba (2010) Interaction of epsteinbarr virus (EBV) with human B-lymphocytes. Biochem. Biophys. Res. Commun. 396: 67-73. https://doi.org/10.1016/j.bbrc.2010.02.146
  6. Durocher, Y. and M. Butler (2009) Expression systems for therapeutic glycoprotein production. Curr. Opin. Biol. 20: 700-707. https://doi.org/10.1016/j.copbio.2009.10.008
  7. van Berkel, P. H. C., J. Gerritsen, E. V. Voskuilen, G. Perdok, T. Vink, J. G. J. van de Winkel, and P. W. H. I. Parren (2010) Rapid production of recombinant human IgG with improved ADCC effector function in a transient expression system. Biotechnol. Bioeng. 105: 350-357. https://doi.org/10.1002/bit.22535
  8. Feng, L. I., X. Joe, X. Yang, T. Tressel, and B. Lee (2005) Current therapeutic antibody production and process optimization. Bioprocessing J. 4: 1-8.
  9. Wurm, F. M. (2004) Production of recombinant protein therapeutics in cultivated mammalian cells. Nat. Biotechnol. 22: 1393-1398. https://doi.org/10.1038/nbt1026
  10. Durrani, A., A. Ali, S. Durrani, J. B. Shaikh, A. Upadhyay, and Z. H. Khan (2011) Non-animal peptone for serum free cultivation of recombinant mammalian and animal cells. Int. J. Biol. 3: 140-145.
  11. Carrier, T., L. Donahue-Hjelle, and M. K. Stramaglia (2009) Banking parental cells according to cGMP guidelines. Bioprocess Int. 12: 20-25.
  12. Pham, P. L., S. Perret, B. Cass, E. Carpentier, G. St-Laurent, L. Bisson, A. Kamen, and Y. Durocher (2005) Transient gene expression in HEK293 cells: peptone addition posttransfection improves recombinant protein synthesis. Biotechnol. Bioeng. 90: 332-344. https://doi.org/10.1002/bit.20428
  13. Sunley, K. and M. Butler (2010) Strategies for the enhancement of recombinant protein production from mammalian cells by growth arrest. Biotechnol. Adv. 28: 385-394. https://doi.org/10.1016/j.biotechadv.2010.02.003
  14. Hosoi, S., H. Miyaji, M. Satoh, T. Kurimoto, A. Mihara, N. Fujiyoshi, S. Itoh, and S. Sato (1991) Optimization of cell culture conditions for production of biologically active proteins. Cytotechnology 5: 17-34. https://doi.org/10.1007/BF00573878
  15. Hosoi, S., K. Murosumi, K. Sasaki, M. Satoh, H. Miyaji, S. Itoh, T. Tamaoki, and S. Sato (1991) Optimization of cell culture conditions for G-CSF (granulocyte-colony stimulating factor) production by genetically engineered Namalwa KJM-1 cells. Cytotechnology 7: 25-32. https://doi.org/10.1007/BF00135635
  16. Heath, C. and R. Kiss (2007) Cell culture process development: advances in process engineering. Biotechnol. Prog. 23: 46-51. https://doi.org/10.1021/bp060344e
  17. Bollin, F., V. Dechavanne, and L. Chevalet (2011) Design of experiment in CHO and HEK transient transfection condition optimization. Protein Expr. Purif. 78: 61-68. https://doi.org/10.1016/j.pep.2011.02.008
  18. Kim, J. Y., Y. G. Kim, Y. K. Han, H. S. Choi, Y. H. Kim, and G. M. Lee (2011) Proteomic understanding of intracellular respones of recombinant chinese hamster ovary cells cultivated in serumfree medium supplemented with hydrolysates. Appl. Microbiol. Biotechnol. 89: 1917-1928. https://doi.org/10.1007/s00253-011-3106-9
  19. Snee, R. D. (2011) Think strategically for design of experiments success. Bioprocess Int. 3: 18-25.
  20. Liu, X., L. Zhao, Y. Wang, X. Zhang, and W. S. Tan (2010) Effects of calcium ion on adenovirus production with high densities of HEK293 cells. Biotechnol. Bioprocess. Eng. 15: 414-420. https://doi.org/10.1007/s12257-009-3032-1
  21. Rajendra, Y., D. Kiseljak, L. Baldi, D. L. Hacker, and F. M. Wurm (2012) Reduced glutamine concentration improves protein production in growth-arrested CHO-DG44 and HEK-293E cells. Biotechnol. Lett. 34: 619-626. https://doi.org/10.1007/s10529-011-0809-z
  22. Shen, C. F. and A. Kamen (2012) Hyperosmotic pressure on HEK 293 cells during the growth phase, but not the production phase, improves adenovirus production. J. Biotechnol. 157: 228-236. https://doi.org/10.1016/j.jbiotec.2011.11.016
  23. Liu, H., X. M. Lou, B. C. Wu, L. L. Ye, X. P. Ni, Q. W. Wang, and Z. L. Chen (2006) Effects of hydrodynamics on aggregates formation, growth and metabolism of HEK 293 cells in suspension culture. Chin. J. Biotechnol. 22: 101-106. https://doi.org/10.1016/S1872-2075(06)60007-1