We are currently trying to co culture mouse primary cortical astrocytes with LUHMES cells. Our initial attempt, where the astrocytes were grown in inserts, before co-culturing with the differentiating LUHMES neurons resulted in the the death of the LUHMES cells. We are currently about to attempt a similar method to the one described by Efremova et al (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4543617/). Does anyone have experience of this and do you have any tips?
Thank you.
Hi! I think luhmes are very sensitive to pH changes. If your astrocytes are very confluent they may be depleting the medium and turn it too acidic. Did you check the color of the medium? I had a similar problem, applied medium from astrocytes (cultured in the luhmes media) to some of my luhmes and when astros were very confluent the luhmes died...we had less confluent astrocytes and luhmes survived and then we realized that pH is really a limiting factor for luhmes..
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