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Writer's pictureSubham

Mammalian Cell Culture: Role of surfactants

#Surfactants play a pivotal role in #mammalian #CellCulture from media to reactor containers to silicone tubings used as connectors. Let us dive deeper to understand what all applications can surfactants have other than being a media component wrt #CellCulture.

Roles

Surfactants serve the following roles in mammalian cell culture:

  • protect cells from shear forces in stirred and bubble aerated environments

  • reduce foaming in stirred/ sparged cultures

  • reduce cell attachment to glass containers, microfluidic devices, silicone tubings etc.

Commonly used surfactants

  • Poloxamer 188 or Pluronic F-68, the most popular one to be used in suspension cell culture medium at a working concentration of 0.1-1%.

Pluronic reduces the interaction between cells and bubbles that arise from sparging by a #decrease in cell-to-bubble attachment, it has a #protective effect on the cells in the bubble-free environment, it is adsorbed on to the membrane surface creating an artificial wall that may protect cells and even #repair damaged (prolonged Trypsin-treated) cells.


There are certain reasons to find and use other surfactants: 1.It can induce #precipitation of concentrated recombinant proteins in certain cases, 2. It begins to #lose effectiveness as the cell count increases, 3. It is highly advisable to have >1 protective surfactant for use in commercial cell culture.

  • Pluronic F-127 is used mostly to coat #PDMS microfluidic devices, #silicone tubings and other surfaces to make them cell-repellent at a working concentration of 0.1-1%. The hydrophobic central part of PF-127 absorbs onto hydrophobic surfaces, presenting the anti-fouling PEG arms away from the surface which actually causes inhibition of cell adhesion.

PF-127 is non-biocidal and can inhibit formation of bacterial biofilms wherever coated, but has lesser anti cell-adhesive properties than PF-68, and can also form unwanted cell aggregates.

  • Maltopyranosides also have similar cytotoxicity levels as Pluronics, are non-crystalline material with high water solubility.

  • Pluronic F-108 has similar properties as the Pluronic Fs, and is largely used to prevent cell adhesion on PDMS.

  • PEG 8000 has shown similar cell growth profiles as PF-68 in T flasks and Shake flasks i.e., small scale experiments at a working concentration of 1% (w/v).

  • PEG 20000 has also shown similar cell growth profiles as PF-68 in T flasks and Shake flasks i.e., small scale experiments at a working concentration of 1% (w/v), slightly beating PEG 8000 in terms of viable cell density after 50 hours incubation.

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