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Related Concept Videos

Fermentation01:29

Fermentation

Most eukaryotic organisms require oxygen to survive and function adequately. Such organisms produce large amounts of energy during aerobic respiration by metabolizing glucose and oxygen into carbon dioxide and water. However, most eukaryotes can generate some energy in the absence of oxygen by anaerobic metabolism.
Fermentation is a type of metabolic process that occurs in the absence of oxygen, where organic molecules such as glucose are broken down to produce energy. During this process, the...
Microbial Fermentation01:23

Microbial Fermentation

Fermentation is a crucial anaerobic metabolic process that enables microbes to derive energy from sugar without relying on oxygen or an electron transport chain. This process is fundamental to various biological and industrial applications and is classified based on the metabolic products generated.Role of Pyruvate in FermentationPyruvate and its derivatives serve as key electron acceptors in fermentative pathways. The oxidation of NADH to regenerate NAD+ is essential for the continuation of...
Bioreactor Design and Operational System01:29

Bioreactor Design and Operational System

Bioreactors are engineered vessels designed to cultivate microorganisms under controlled conditions for industrial bioprocessing. They maintain sterility and allow precise regulation of pH, temperature, oxygen, and nutrient levels to optimize microbial growth and metabolite production. Bioreactors range from small laboratory units of 1 liter to industrial systems holding up to 500,000 liters, though only about 75% of their volume is actively used for fermentation. The remaining headspace...
Bioreactor Controls-I01:28

Bioreactor Controls-I

Maintaining optimal conditions within fermenters is essential for maximizing microbial productivity and ensuring process efficiency. This lesson focuses on key parameters—temperature, foam, pH, carbon dioxide, oxygen, and pressure—and their precise measurement and control strategies in fermentation systems.Temperature ControlTemperature regulation is critical due to the exothermic nature of many fermentation processes. In small laboratory fermenters, temperature is commonly monitored using...
Bioreactor Controls-II01:18

Bioreactor Controls-II

In aerobic fermentations, oxygen is vital for microbial growth and metabolite production. Since air comprises only about 20% oxygen and the gas is poorly soluble in water—just 9 ppm at 20°C—supplying sufficient oxygen becomes a critical challenge, especially in high-demand processes like yeast growth or citric acid production. Even a fully saturated broth may offer only a few seconds of oxygen availability.To address this, sterile or scrubbed air is introduced into the fermentor via a sparger...
Scale-Up Processes01:14

Scale-Up Processes

The scale-up of microbial fermentation processes is essential in industrial biotechnology, allowing the transition from laboratory-scale experiments to commercial-scale production while aiming to maintain product yield and quality. This process requires meticulous adjustment of equipment design, process parameters, and contamination control strategies to accommodate increasing culture volumes.At the laboratory scale, cultures are typically maintained in 1 to 10-liter glass or autoclavable...

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Related Experiment Video

Updated: Jun 22, 2026

Scale-Up of Mammalian Cell Culture using a New Multilayered Flask
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Reinventing shake flask fermentation: The breathable flask.

Vikash Kumar1,2, Michael Tolosa2, Xudong Ge1,2

  • 1Department of Chemical, Biochemical and Environmental Engineering, University of Maryland, Baltimore County, Baltimore, USA.

Biotechnology and Bioengineering
|May 3, 2024
PubMed
Summary

This study introduces a breathable flask that enhances cell culture by improving gas exchange. The novel design boosts biomass and product yield in both prokaryotic and eukaryotic cell cultures.

Keywords:
breathablecell culturefermentationmass transfershake flask

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Area of Science:

  • Biotechnology
  • Bioprocess Engineering
  • Cell Culture Technology

Background:

  • Conventional shake flask cultures face limitations in gas exchange, leading to oxygen deprivation and carbon dioxide buildup.
  • These mass transfer issues significantly impact cell growth and recombinant protein production.
  • Current methods for monitoring cell cultures in shake flasks are often invasive or lack real-time data.

Purpose of the Study:

  • To develop and evaluate a novel shake flask design that overcomes traditional mass transfer limitations.
  • To enhance oxygen and carbon dioxide exchange between the cell culture and the external environment.
  • To create a platform for integrated, non-invasive sensing of critical cell culture parameters.

Main Methods:

  • Modification of a conventional shake flask by replacing the vessel wall with a permeable membrane to create a 'breathable flask'.
  • Determination of the mass transfer coefficient (kLa) using a static diffusion method.
  • Conducting prokaryotic (Escherichia coli) and eukaryotic (Pichia pastoris) cell cultures to assess performance.
  • Evaluating biomass and recombinant product yield improvements.

Main Results:

  • The breathable flask demonstrated a 40% improvement in the mass transfer coefficient (kLa).
  • Prokaryotic cultures showed 28%-66% increase in biomass and 41%-56% increase in product yield.
  • Eukaryotic cultures exhibited a 40% increase in biomass and a significant 115% increase in protein yield.
  • The membrane-diffusion design enables potential for integrated real-time sensing of dissolved oxygen and carbon dioxide.

Conclusions:

  • The breathable flask effectively addresses mass transfer limitations inherent in conventional shake flask cultivation.
  • This innovative design significantly enhances both cell biomass and recombinant product yields.
  • The integrated sensing capability offers a promising avenue for advanced, non-invasive monitoring of cell cultures.