Product FAQ

Why does the Insect cell line manual state: "Cells should be maintained at 27°C in a non-humidified environment."

Answer

Insect cells do not require CO2 or high humidity to grow, they can grow in a lab drawer at room temperature. We recommend this so people don't waste CO2 and other resources necessary for maintaining a tissue culture incubator. It should be noted, however, that the cells will grow in a humidified incubator.

Answer Id: E3309

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Citations & References

Endogenous K+/H+ exchange activity in the Sf9 insect cell line.

  • Authors: Vachon V, Paradis MJ, Marsolais M, Schwartz JL, Laprade R
  • Journal: Biochemistry (1995) 34:15157-15164
  • PubMed ID: 7578130
Catalog #

Citations & References

Baculovirus immediate-early promoter-mediated expression of the Zeocin resistance gene for use as a dominant selectable marker in dipteran and lepidopteran insect cell lines.

  • Authors: Pfeifer TA, Hegedus DD, Grigliatti TA, Theilmann DA,
  • Journal: Gene (1997) 188:183-190
  • PubMed ID: 9133590

Citations & References

Tnao38, high five and Sf9--evaluation of host-virus interactions in three different insect cell lines: baculovirus production and recombinant protein expression.

  • Authors: Wilde M, Klausberger M, Palmberger D, Ernst W, Grabherr R
  • Journal: Biotechnol Lett 2014; (36):4 743-749
  • PubMed ID: 24375231
Catalog #

Citations & References

Equilibrium and kinetic analysis of Autographa californica nuclear polyhedrosis virus attachment to different insect cell lines.

  • Authors: Wickham TJ, Shuler ML, Hammer DA, Granados RR, Wood HA
  • Journal: J Gen Virol 1992 (73 ( Pt 12)):3185-3194
  • PubMed ID: 1469356

Manual / Product Insert

User Guide: Growth and Maintenance of Insect Cell Lines

Version: MAN0000030 Rev B.0 (14Mar2017)

Citations & References

In vitro infection of Wolbachia in insect cell lines

  • Authors: Furukawa, S; Tanaka, K; Fukatsu, T; Sasaki, T
  • Journal: APPLIED ENTOMOLOGY AND ZOOLOGY 2008 4:519-525

Product FAQ

What are the doubling times for robust growth and maintenance of different insect cell lines?

Answer

Please see below for information related to the type of cells, media, and doubling time in hours:

D.mel-2; Schneider’s Drosophila + 10% FBS heat-inactivated; 18 to 24
High Five cells; Express Five SFM; approximately 24
Sf9 cells; Sf-900 II SFM; 24 to 30
Sf9 cells; Sf-900 III SFM; 24 to 30
Sf21 cells; Sf-900 II SFM; 24 to 30
Sf21 cells Sf-900 III SFM; 24 to 30

Answer Id: E11932

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Product FAQ

Can I adapt my own insect cell line for growth in ExpiSf CD Medium?

Answer

Yes, you may be able to adapt your Sf9 cells for growth in ExpiSf CD Medium. We recommend following a serial adaptation protocol to slowly acclimate your cells for growth in ExpiSf CD Medium. A detailed cell adaptation protocol is provided in the ExpiSf CD Medium User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017561_ExpiSfExpressSystem_SmartStart_UG.pdf). Long-term adaptation in ExpiSf CD Medium may increase productivity of your Sf9 cells, and should support high-density growth. However, there is no guarantee that other insect cell lines will achieve the same levels of baculovirus and protein expression as the ExpiSf9 Cells. In limited testing, we have found other Sf9 and Sf21 cell lines to achieve lower maximum cell densities when fully adapted to ExpiSf CD Medium with lower protein yields. The performance of your own insect cell line in ExpiSf CD Medium will need to be empirically determined.

Answer Id: E16211

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Citations & References

Screening of insect cell lines for the production of recombinant proteins and infectious virus in the baculovirus expression system.

  • Authors: Wickham TJ, Davis T, Granados RR, Shuler ML, Wood HA
  • Journal: Biotechnol Prog (8):391-396
  • PubMed ID: 1369220

Citations & References

Latent infection of a new alphanodavirus in an insect cell line.

  • Authors: Li TC; Scotti PD; Miyamura T; Takeda N
  • Journal: Journal of Virology 2008 20:10890-10896

Citations & References

Comparative recombinant protein production of eight insect cell lines.

  • Authors: Davis TR, Wickham TJ, McKenna KA, Granados RR, Shuler ML, Wood HA
  • Journal: In Vitro Cell Dev Biol Anim 1993 (29A):388-390
  • PubMed ID: 8314732

Product FAQ

Which insect cell line is best to use?

Answer

We recommend Sf9 or Sf21 cells for transfection, purification, and amplification of recombinant virus. Sf9 cells are regular in size, easy to manipulate, and form good monolayers for plaque assays. Sf9 and Sf21 cells can also be used for expression of recombinant proteins, but the High Five cell line may produce higher levels. We recommend the High Five cell line for expression of secreted recombinant proteins. They are grown in serum-free medium, adaptable to suspension culture, and produce high levels of recombinant protein (Davis et al., 1992; see http://www.ncbi.nlm.nih.gov/pubmed/1368794).

Note: Generally it is easier to use one cell line for procedures up to optimization of protein expression. Once you have confirmed expression of your recombinant protein, other cell lines can be tried for optimization of expression levels.

Answer Id: E11936

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Product FAQ

I don't have ExpiSf9 Cells. Can ExpiFectamine Sf Transfection Reagent be used to transfect other insect cells?

Answer

Yes. The ExpiFectamine Sf Transfection Reagent can be used to efficiently transfect Sf9 and Sf21 cells. The reagent can be used to transfect these cells in both adherent and suspension culture formats.

Answer Id: E16217

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Product FAQ

What are the normal growth characteristics of ExpiSf9 Cells?

Answer

ExpiSf9 cells should exhibit ≥80% viability at 3 days post-thaw. Within 1-2 passages post-thaw, the cells should be growing with a doubling time of approximately 24 hours. When cells are cultured at 0.5 x 10E6 cells/mL, or 1.0 x 10E6 cells/mL, viable cell density should be approximately 5-10 x 10E6 cells/mL within 3 or 4 days, respectively. If cells are not growing within these approximate ranges within 3 passages from thawing, cell culture conditions will require further optimization.

Answer Id: E16192

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