TubeSpin bioreactor 50 for the high-density cultivation of Sf-9 insect cells in suspension (original) (raw)

Abstract

Here we present the TubeSpin bioreactor 50 (TubeSpins) as a simple and disposable culture system for Sf-9 insect cells in suspension. Sf-9 cells had substantially better growth in TubeSpins than in spinner flasks. After inoculation with 106 cells/ml, maximal cell densities of 16 × 106 and 6 × 106 cells/ml were reached in TubeSpins and spinner flasks, respectively. In addition the cell viability in these batch cultures remained above 90% for 10 days in TubeSpins but only for 4 days in spinner flasks. Inoculation at even higher cell densities reduced the duration of the lag phase. After inoculation at 2.5 × 106 cells/ml, the culture reached the maximum cell density within 3 days instead of 7 days as observed for inoculation with 106 cells/ml. Infection of Sf-9 cells in TubeSpins or spinner flasks with a recombinant baculovirus coding for green fluorescent protein (GFP) resulted in similar GFP-specific fluorescence levels. TubeSpins are thus an attractive option for the small-scale cultivation of Sf-9 cells in suspension and for baculovirus-mediated recombinant protein production.

Access this article

Log in via an institution

Subscribe and save

• Starting from 10 chapters or articles per month
• Access and download chapters and articles from more than 300k books and 2,500 journals
• Cancel anytime View plans

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1

The alternative text for this image may have been generated using AI.

Fig. 2

The alternative text for this image may have been generated using AI.

Fig. 3

The alternative text for this image may have been generated using AI.

Fig. 4

The alternative text for this image may have been generated using AI.

Similar content being viewed by others

References

• Batista FR, Pereira CA, Mendonça RZ, Moraes AM (2005) Enhancement of Sf9 cells and baculovirus production employing Grace’s medium supplemented with milk whey ultrafiltrate. Cytotechnology 49(1):1–9
  Article PubMed CAS Google Scholar
• Bédard C, Tom R, Kamen A (1993) Growth, nutrient consumption, and end product accumulation in Sf-9 and BTI-EAA insect cell cultures: insights into growth limitation and metabolism. Biotechnol Prog 9(6):615–621
  Article PubMed Google Scholar
• Bédard C, Perret S, Kamen AA (1997) Fed-batch culture of Sf-9 cells supports 3 × 107 cells/ml and improves baculovirus-expressed recombinant protein yields. Biotechnol Lett 19:629–632
  Article Google Scholar
• Berger I, Fitzgerald DJ, Richmond TJ (2004) Baculovirus expression system for heterologous multiprotein complexes. Nat Biotechnol 22(12):1583–1587
  Article PubMed CAS Google Scholar
• Bovo R, Galesi AL, Jorge SA, Piccoli RA, Moraes AM, Pereira CA, Augusto EF (2008) Kinetic response of a Drosophila melanogaster cell line to different medium formulations and culture conditions. Cytotechnology 57(1):23–35
  Article PubMed CAS Google Scholar
• Chiou T-W, Hsieh Y-C, Ho CS (2000) High density culture of insect cells using rational medium design and feeding strategy. Bioprocess Biosyst Eng 22(6):483–491
  CAS Google Scholar
• DeJesus MJ, Girard P, Bourgeois M, Baumgartner G, Jacko B, Amstutz H, Wurm FM (2004) TubeSpin satellites: a fast track approach for process development with animal cells using shaking technology. Biochem Eng J 17:217–223
  Article CAS Google Scholar
• Deparis V, Durrieu C, Schweizer M, Goergen JL, Chevalot I, Marc A (2003) Promoting effect of rapeseed proteins and peptides on Sf9 insect cell growth. Cytotechnology 42(2):75–85
  Article PubMed CAS Google Scholar
• Diao J, Young L, Zhou P, Shuler ML (2008) An actively mixed mini-bioreactor for protein production from suspended animal cells. Biotechnol Bioeng 100:72–81
  Article PubMed CAS Google Scholar
• Drews M, Paalme T, Vilu R (1995) The growth and nutrient utilization of the insect cell line Spodoptera frugiperda Sf9 in batch and continuous culture. J Biotechnol 40:187–198
  Article CAS Google Scholar
• Elias CB, Zeiser A, Bédard C, Kamen AA (2000) Enhanced growth of Sf-9 cells to a maximum density of 5.2 × 10(7) cells per ml and production of beta-galactosidase at high cell density by fed batch culture. Biotechnol Bioeng 68(4):381–388
  Article PubMed CAS Google Scholar
• Gotoh T, Chiba K, Kikuchi K-I (2004) Oxygen consumption profiles of Sf-9 insect cells and their culture at low temperature to circumvent oxygen starvation. Biochem Eng J 17(2):71–78
  Article Google Scholar
• Grace TD (1962) Establishment of four strains of cells from insect tissues grown in vitro. Nature 195:788–789
  Article PubMed CAS Google Scholar
• Ikonomou L, Schneider Y-J, Agathos SN (2003) Insect cell culture for industrial production of recombinant proteins. Appl Microbiol Biotechnol 62(1):1–20
  Article PubMed CAS Google Scholar
• Jäger V (1996) Perfusion bioreactors for the production of recombinant proteins in insect cells. Cytotechnology 20:191–198
  Article Google Scholar
• Kost TA, Condreay JP (1999) Recombinant baculoviruses as expression vectors for insect and mammalian cells. Curr Opin Biotechnol 10(5):428–433
  Article PubMed CAS Google Scholar
• Kost TA, Condreay JP, Jarvis DL (2005) Baculovirus as versatile vectors for protein expression in insect and mammalian cells. Nat Biotechnol 23(5):567–575
  Article PubMed CAS Google Scholar
• Marheineke K, Gruënewald S, Christie W, Reiländer H (1998) Lipid composition of Spodoptera frugiperda (Sf9) and Trichoplusia ni (Tn) insect cells used for baculovirus infection. FEBS Lett 441(1):49–52
  Article PubMed CAS Google Scholar
• Muller N, Girard P, Hacker DL, Jordan M, Wurm FM (2005) Orbital shaker technology for the cultivation of mammalian cells in suspension. Biotechnol Bioeng 89(4):400–406
  Article PubMed CAS Google Scholar
• Nienow AW (2006) Reactor engineering in large scale animal cell culture. Cytotechnology 50(1–3):9–33
  Article PubMed CAS Google Scholar
• Öhman L, Alarcon M, Ljunggren J, Ramqvist A-K, Häggström L (1996) Glutamine is not an essential amino acid for Sf-9 insect cells. Biotechnol Lett 18(7):765–770
  Article Google Scholar
• Ojala K, Mottershead DG, Suokko A, Oker-Blom C (2001) Specific binding of baculoviruses displaying gp64 fusion proteins to mammalian cells. Biochem Biophys Res Commun 284:777–784
  Article PubMed CAS Google Scholar
• Oker-Blom C, Orellana A, Keinanen K (1996) Highly efficient production of GFP and its derivatives in insect cells for visual in vitro applications. FEBS Lett 389:238–243
  Article PubMed CAS Google Scholar
• Zhang X, Bürki C-A, Stettler M, De Sanctis D, Perrone M, Discacciati M, Parolini N, DeJesus M, Hacker DL, Quarteroni A, Wurm FM (2009) Efficient oxygen transfer by surface aeration in shaken cylindrical containers for mammalian cell cultivation at volumetric scales up to 1000 L. Biochem Eng J 45:41–47
  Article Google Scholar

Download references

Acknowledgments

The authors thank Dr. Hilal Lashuel of the EPFL for the Sf-9 cells and Dr. Leona Gilbert of the University of Jyväskylä for the recombinant baculovirus. This study was supported by grants from the Guangdong Provincial Department of Science and Technology (2008B030301349), the MOE of China (211 Grant), and the Academy of Finland (decision no. 135820).

Author information

Author notes

1. Xiaowei Zhang
   Present address: College of Translational Medicine, Nanchang University, Nanchang, 330031, Jiangxi, People’s Republic of China

Authors and Affiliations

1. Laboratory of Cellular Biotechnology, Institute of Bioengineering, École Polytechnique Fédérale de Lausanne, CH J2 506, Station 6, 1015, Lausanne, Switzerland
   Qiuling Xie, Patrik O. Michel, Lucia Baldi, David L. Hacker, Xiaowei Zhang & Florian M. Wurm
2. Institute of Bioengineering, Jinan University, Guangzhou, 510632, Guangdong, People’s Republic of China
   Qiuling Xie

Authors

1. Qiuling Xie
2. Patrik O. Michel
3. Lucia Baldi
4. David L. Hacker
5. Xiaowei Zhang
6. Florian M. Wurm

Corresponding author

Correspondence toFlorian M. Wurm.

Rights and permissions

About this article

Cite this article

Xie, Q., Michel, P.O., Baldi, L. et al. TubeSpin bioreactor 50 for the high-density cultivation of Sf-9 insect cells in suspension.Biotechnol Lett 33, 897–902 (2011). https://doi.org/10.1007/s10529-011-0527-6

Download citation

• Received: 20 August 2010
• Accepted: 23 December 2010
• Published: 26 January 2011
• Issue date: May 2011
• DOI: https://doi.org/10.1007/s10529-011-0527-6

Keywords