BrainPhys neuronal medium optimized for imaging and optogenetics in vitro

Michael Zabolocki, Kasandra McCormack, Mark Hurk, Bridget Milky, Andrew P. Shoubridge, Robert Adams, Jenne Tran, Anita Mahadevan-Jansen, Philipp Reineck, Jacob Thomas, Mark R. Hutchinson, Carmen K. H. Mak, Adam Añonuevo, Leon H. Chew, Adam J. Hirst, Vivian M. Lee, Erin Knock and Cedric Bardy ()
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Michael Zabolocki: South Australian Health and Medical Research Institute (SAHMRI)
Kasandra McCormack: STEMCELL Technologies
Mark Hurk: South Australian Health and Medical Research Institute (SAHMRI)
Bridget Milky: South Australian Health and Medical Research Institute (SAHMRI)
Andrew P. Shoubridge: South Australian Health and Medical Research Institute (SAHMRI)
Robert Adams: South Australian Health and Medical Research Institute (SAHMRI)
Jenne Tran: South Australian Health and Medical Research Institute (SAHMRI)
Anita Mahadevan-Jansen: Vanderbilt University
Philipp Reineck: RMIT University
Jacob Thomas: University of Adelaide
Mark R. Hutchinson: University of Adelaide
Carmen K. H. Mak: STEMCELL Technologies
Adam Añonuevo: STEMCELL Technologies
Leon H. Chew: STEMCELL Technologies
Adam J. Hirst: STEMCELL Technologies
Vivian M. Lee: STEMCELL Technologies
Erin Knock: STEMCELL Technologies
Cedric Bardy: South Australian Health and Medical Research Institute (SAHMRI)

Nature Communications, 2020, vol. 11, issue 1, 1-19

Abstract: Abstract The capabilities of imaging technologies, fluorescent sensors, and optogenetics tools for cell biology are advancing. In parallel, cellular reprogramming and organoid engineering are expanding the use of human neuronal models in vitro. This creates an increasing need for tissue culture conditions better adapted to live-cell imaging. Here, we identify multiple caveats of traditional media when used for live imaging and functional assays on neuronal cultures (i.e., suboptimal fluorescence signals, phototoxicity, and unphysiological neuronal activity). To overcome these issues, we develop a neuromedium called BrainPhys™ Imaging (BPI) in which we optimize the concentrations of fluorescent and phototoxic compounds. BPI is based on the formulation of the original BrainPhys medium. We benchmark available neuronal media and show that BPI enhances fluorescence signals, reduces phototoxicity and optimally supports the electrical and synaptic activity of neurons in culture. We also show the superior capacity of BPI for optogenetics and calcium imaging of human neurons. Altogether, our study shows that BPI improves the quality of a wide range of fluorescence imaging applications with live neurons in vitro while supporting optimal neuronal viability and function.

Date: 2020
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DOI: 10.1038/s41467-020-19275-x

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