, Volume 68, Issue 6, pp 2649–2658 | Cite as

Carbohydrates–chitosan composite carrier for Vero cell culture

  • Ya-Ching Lin
  • Guan-Ting Chen
  • Sheng-Chi Wu
Original Article


In this study, carbohydrate–chitosan composite including glucose–chitosan, sucrose–chitosan and starch–chitosan with varied carbohydrate concentrations were prepared as carriers for Vero cell culture. Our results show that among these composites, 30 % starch–chitosan composite (STC) were the best carriers for the growth of Vero cells. The initial number of attached cells on the surface of composite carriers did not have any significant effect on subsequent cell production. A higher glucose level in the growth medium during the exponential phase of cell growth, however, played an important factor for cell production. Vero cells on the STC carriers were able to convert starch inside the composite carriers into glucose and further utilized the glucose for their growth. Moreover, by crosslink with serum the STC carriers supported an even better cell production in the normal medium without adding fetal bovine serum, as well as a good extracellular virus production. The STC composite is therefore a promising alternative carrier for Vero cell culture.


Cell carrier Carbohydrates Chitosan Vero cell 


  1. Adina C, Fetea F, Taoutaou A, Socaciu C (2010) Application of FTIR spectroscopy for a rapid determination of some hydrolytic enzymes activity on sea buckthorn substrate. Rom Biotechnol Lett 15:5738–5744Google Scholar
  2. Alberta Araújo M, Cunha AM, Mota M (2004) Enzymatic degradation of starch-based thermoplastic compounds used in protheses: identification of the degradation products in solution. Biomaterials 25:2687–2693CrossRefGoogle Scholar
  3. Bačáková L, Novotná K, Pařízek M (2014) Polysaccharides as cell carriers for tissue engineering: the use of cellulose in vascular wall reconstruction. Physiol Res 63:22Google Scholar
  4. Baran ET, Mano JF, Reis RL (2004) Starch–chitosan hydrogels prepared by reductive alkylation cross-linking. J Mater Sci Mater Med 15:759–765CrossRefGoogle Scholar
  5. Baran ET, Tuzlakoglu K, Mano JF, Reis RL (2012) Enzymatic degradation behavior and cytocompatibility of silk fibroin–starch–chitosan conjugate membranes. Mater Sci Eng C Mater Biol Appl 32:1314–1322CrossRefGoogle Scholar
  6. Brunner D, Frank J, Appl H, Schöffl H, Pfaller W, Gstraunthaler G (2010) Serum-free cell culture: the serum-free media interactive online database. Altex 27:53–62Google Scholar
  7. da Costa-Silva TA, da Silva Meira C, Frazzatti-Gallina N, Pereira-Chioccola VL (2012) Toxoplasma gondii antigens: recovery analysis of tachyzoites cultivated in Vero cell maintained in serum free medium. Exp Parasitol 130:463–469CrossRefGoogle Scholar
  8. Gao W, Lai JC, Leung SW (2012) Functional enhancement of chitosan and nanoparticles in cell culture, tissue engineering, and pharmaceutical applications. Front Physiol 3:321CrossRefGoogle Scholar
  9. Han Y, Liu XM, Liu H, Li SC, Wu BC, Ye LL, Wang QW, Chen ZL (2006) Cultivation of recombinant Chinese hamster ovary cells grown as suspended aggregates in stirred vessels. J Biosci Bioeng 102:430–435CrossRefGoogle Scholar
  10. Huang CB, Robert J, Mohini S, Bradley A, Martin H (2006) Production, characterization, and mechanical properities of starch modified by Ophiostoma Spp. BioResources 1:257–269Google Scholar
  11. Kishimoto S, Nakamura S, Nakamura S, Kanatani Y, Hattori H, Tanaka Y, Harada Y, Tagawa M, Mori Y, Maehara T, Ishihara M (2009) Fragmin/protamine microparticle-coated matrix immobilized cytokines to stimulate various cell proliferations with low serum media. Artif Organs 33:431–438CrossRefGoogle Scholar
  12. Li J, Pan J, Zhang L, Yu Y (2003) Culture of hepatocytes on fructose-modified chitosan scaffolds. Biomaterials 24:2317–2322CrossRefGoogle Scholar
  13. Lu DR (2009) Starch-based completely biodegradable polymer materials. Express Polym Lett 3:366–375CrossRefGoogle Scholar
  14. Luangbudnark W, Viyoch J, Laupattarakasem W, Surakunprapha P, Laupattarakasem P (2012) Properties andbiocompatibility of chitosan and silk fibroin blend films for application in skin tissue engineering. Sci World J 2012:697201CrossRefGoogle Scholar
  15. Maitra J, Singh N (2014) Swelling behavior of starch chitosan polymeric blend. Adv Polym Sci Technol Int J 4:22–27Google Scholar
  16. Mattila PK, Lappalainen P (2008) Filopodia: molecular architecture and cellular functions. Nat Rev Mol Cell Biol 9:446–454CrossRefGoogle Scholar
  17. Medhat I, Moussa A, Hanan E, Abraham FJ, Aned DL (2006) Analysis of the structure and vibrational spectra of glucose and fructose. Eclet Quim 31:15–21CrossRefGoogle Scholar
  18. Merten OW, Cruz PE, Rochette C, Geny-Fiamma C, Bouquet C, Gonçalves D, Danos O, Carrondo MJ (2001) Comparison of different bioreactor systems for the production of high titer retroviral vectors. Biotechnol Prog 17:326–335CrossRefGoogle Scholar
  19. Nienow AW (2006) Reactor engineering in large scale animal cell culture. Cytotechnology 50:9–33CrossRefGoogle Scholar
  20. Reid LM (1990) Stem cell biology, hormone/matrix synergies and liver differentiation. Curr Opin Cell Biol 2:121–130CrossRefGoogle Scholar
  21. Rodrigues AI, Gomes ME, Leonor IB, Reis RL (2012) Bioactive starch-based scaffolds and human adipose stem cells are a good combination for bone tissue engineering. Acta Biomater 8:3765–3776CrossRefGoogle Scholar
  22. Silva SS, Santos M, Coutinho O, Mano J, Reis R (2005) Physical properties and biocompatibility of chitosan/soy blended membranes. J Mater Sci Mater Med 16:575–579CrossRefGoogle Scholar
  23. Silva SS, Mano JF, Reis RL (2010) Potential applications of natural origin polymer-based systems in soft tissue regeneration. Crit Rev Biotechnol 30:200–221CrossRefGoogle Scholar
  24. Silva SS, Caridade SG, Mano JF, Reis RL (2013) Effect of crosslinking in chitosan/aloe vera-based membranes for biomedical applications. Carbohydr Polym 98:581–588CrossRefGoogle Scholar
  25. Singh N, Maitra J (2015) Antibacterial evaluation of starch and chitosan based polymeric blend. IOSR J Appl Chem 8:26–32Google Scholar
  26. Subramanian SB, Francis AP, Devasena T (2014) Chitosan-starch nanocomposite particles as a drug carrier for the delivery of bis-desmethoxy curcumin analog. Carbohydr Polym 114:170–178CrossRefGoogle Scholar
  27. Toriniwa H, Komiya T (2008) Long-term stability of Vero cell-derived inactivated Japanese encephalitis vaccine prepared using serum-free medium. Vaccine 26:3680–3689CrossRefGoogle Scholar
  28. Trabelsi K, Rourou S, Loukil H, Majoul S, Kallel H (2006) Optimization of virus yield as a strategy to improve rabies vaccine production by Vero cells in a bioreactor. J Biotechnol 121:261–271CrossRefGoogle Scholar
  29. van der Loo JC, Swaney WP, Grassman E, Terwilliger A, Higashimoto T, Schambach A, Baum C, Thrasher AJ, Williams DA, Nordling DL, Reeves L, Malik P (2012) Scale-up and manufacturing of clinical-grade self-inactivating gamma-retroviral vectors by transient transfection. Gene Ther 19:246–254CrossRefGoogle Scholar
  30. Wang C, Yang F, Meng F, Zhang H, Xue Y, Fu G (2010) High flux and antifouling filtration membrane based on non-woven fabric with chitosan coating for membrane bioreactors. Bioresour Technol 101:5469–5474CrossRefGoogle Scholar
  31. Wu SC, Liu CC, Lian WC (2004) Optimization of microcarrier cell culture process for the inactivated enterovirus type 71 vaccine development. Vaccine 22:3858–3864CrossRefGoogle Scholar
  32. Zhang H, Wang W, Quan C, Fan S (2010) Engineering considerations for process development in mammalian cell cultivation. Curr Pharm Biotechnol 11:103–112CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  1. 1.Department of BiotechnologyFooyin UniversityKaohsiung CityTaiwan

Personalised recommendations