Effect of lipids on insect cell growth and expression of recombinant proteins in serum-free medium (original) (raw)

Abstract

The lipid emulsion components of a serum-free insect cell medium were varied and evaluated for effects on cell growth and recombinant protein expression. The growth of High-FiveTM cells was significantly affected by polyol Pluronic F-68 and Tween-80, but not by lipids. Pluronic was essential for cell growth, while Tween-80 was required to achieve maximum cell densities. A dose response effect was observed for Tween-80 with optimal cell growth at a concentration of 25 mg/l. Cholesterol had a minor effect on cell growth, but was essential for the expression of recombinant proteins. The expression of β-galactosidase (β-gal) was directly affected by cholesterol with optimal expression at a concentration of 5.4 mg/l. Vitamin E, important as an antioxidant to stabilize lipids, did not directly affect recombinant protein expression. Although lipids were not required for cell growth, the presence of lipids were required during the cell growth phase in order to achieve efficient infection with baculovirus. These studies help to define the important components, and range of concentrations, for lipid emulsions which can effectively replace serum in insect cell culture.

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.

Similar content being viewed by others

Abbreviations

β-gal:

galactosidase (E.C. 3.2.1.23)

Sf-9:

Spodoptera frugiperda

High-5:

Trichoplusia ni 5Bl-4

References

• Bignon C, Daniel N and Djiane J (1993) β-Galactosidase and chloramphenicol acetyltransferase assays in 96-well plates. Biotechniques 15: 243–246.
  Google Scholar
• Davis TR, Wickham TJ, McKenna KA, Granados RR, Shuler ML and Wood HA (1993) Comparative recombinant protein production of eight insect cell lines. In Vitro Cell Dev Biol 29A: 388–390.
  Google Scholar
• Goodwin RH (1991) Replacement of vertebrate serum with lipids and other factors in the culture of invertebrate cells, tissues, parasites, and pathogens. In Vitro Cell Dev Biol 27A: 470–478.
  Google Scholar
• Hink WF (1991) A serum-free medium for the cultivation of insect cells and production of recombinant proteins. In Vitro Cell Dev Biol 29A: 397–401.
  Google Scholar
• Luckow VA (1991) Cloning and expression of heterologous genes in insect cells with baculovirus vectors. In: Ho C, Prokop A, Baipai R (eds) Recombinant DNA technology and applications (pp. 97–152) McGraw-Hill, New York.
  Google Scholar
• Maiorella B, Inlow D and Shauger A, et al. (1988) Large-scale insect cell-culture for recombinant protein production. Bio/Technology 6: 1406–1410.
  Google Scholar
• Miller LK (1993) Baculoviruses: High-level expression in insect cells. Curr Opin Genet Dev 3: 97–101.
  Google Scholar
• Mitsuhashi J (1989) Nutritional requirements of insect cells in vitro. In: Invertebrate cell system applications. Vol. 1 (pp. 3–20, review) CRC Press, Florida.
  Google Scholar
• Murhammer DW and Goochee CF (1988) Scaleup of insect cell cultures: protective effects of Pluronic F-68. Bio/Technology 6: 1411–1418.
  Google Scholar
• Roeder A (1982) Development of a serum-free medium for cultivation of insect cells. Naturwissenschaften 69: 92–93.
  Google Scholar

Download references

Author information

Authors and Affiliations

1. Irvine Scientific, 2511 Daimler St., 92705-5588, Santa Ana, CA, USA
   Rebecca S. Gilbert, Yuichi Nagano, Tadafumi Yokota, Sun Fon Hwan, Tom Fletcher & Ken Lydersen

Authors

1. Rebecca S. Gilbert
2. Yuichi Nagano
3. Tadafumi Yokota
4. Sun Fon Hwan
5. Tom Fletcher
6. Ken Lydersen

Rights and permissions

About this article

Cite this article

Gilbert, R.S., Nagano, Y., Yokota, T. et al. Effect of lipids on insect cell growth and expression of recombinant proteins in serum-free medium.Cytotechnology 22, 211–216 (1996). https://doi.org/10.1007/BF00353941

Download citation

• Issue date: January 1996
• DOI: https://doi.org/10.1007/BF00353941

Key words