Abstract

Growing insect cells with high air space in culture vessel is common from the early development of suspension cell culture. We believed and followed it with the hope that it allows sufficient air for optimal cell growth. However, we missed to identify how much air exactly cells need for its growth and multiplication. Here we present the innovative method that changed the way we run insect cell culture. The method is easy to adapt, cost-effective and useful for both academic and industrial research labs. We believe this method will revolutionize the way we run insect cell culture by increasing throughput in a cost-effective way.In our study we identified:•Insect cells need to be in suspension; air space in culture vessel and type of culture vessel is of less importance. Shaking condition that introduces small air bubbles and maintains it in suspension for longer time provides better oxygen transfer in liquid. For this, high-fill volume in combination with speed and shaking diameter are important.•Commercially available insect cells are not fragile as original isolates. These cells can easily withstand higher shaking speed.•Growth condition in particular lab set-up needs to be optimized. The condition used in one lab may not be optimum for another lab due to different incubators from different vendors.

Highlights

  • Shaking bioreactors are widely used in academia and in bio-industry for screening and bioprocess development [1]

  • Growth of insect cells in reagent bottle serves better alternative to carry out multi-parallel cultures in volume ranges from 0.25 L to 5 L

  • Sf9 or Sf21 cells adapted to suspension culture in Sf-900TM III SFM medium (Cat. no. 12658-027, life technologies) were grown to log phase, i.e. maintaining viable count in the range of (2–5) Â 106 cells per ml and viability greater than 95%

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Summary

GRAPHICAL ABSTRACT

ABSTRACT Growing insect cells with high air space in culture vessel is common from the early development of suspension cell culture. The method is easy to adapt, costeffective and useful for both academic and industrial research labs. We believe this method will revolutionize the way we run insect cell culture by increasing throughput in a cost-effective way. In our study we identified: Insect cells need to be in suspension; air space in culture vessel and type of culture vessel is of less importance. Available insect cells are not fragile as original isolates. These cells can withstand higher shaking speed. ARTICLE INFO Method: Insect cell culture in reagent bottles Keywords: Insect cell, Baculovirus, BV, Sf9, Sf21, High-throughput, Fermentation vessel, Reagent bottle, High volume culture, Expression Article history: Received 9 July 2014; Accepted 13 August 2014; Available online 28 August 2014

Preparation of reagent bottles for cell culture
Routine growth and maintenance of cells
Shaking diameter
Growth behavior over different passages
Background
BoƩle SpinTube
Findings
Cell line
Full Text
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