I have been having difficulties in trying to culture mouse DRG neurons in microfluidic devices, that help isolate neuronal cell body from axons (bought from Xona SND150). My problem is with getting the dissociated DRG neurons to attach to the main channel of the device. I coat the device overnight with 0.5 mg/ml PDL followed by 100 ug/ml Laminin for 4-5 hours. I quickly rinse the device 3 times after each coating. I flow in 100,000 cells in a small volume (18 ul) slowly into the main channel (10 ul from the top well and 8 ul from the bottom well to dampen the flow through the main channel). Even though I get a lot of cells to stay in the main channel at this point, most of these don't seem to attach - they slowly drift away either during the 1 hr period I give for the cells to attach or when I add in more medium after that into the device.
Would appreciate any suggestions and thanks for your help!
I have no experience with microfluidic devices but I have found that the best attachment for neurons is using PEI (polyethyleneimine) but it must be dried overnight before applying the cells.
Thanks, Elizabeth. I will try using PEI and see if that works better.
Hi, we have successfully grown DRG neurons using Ananda Microfluidic Devices, here is the instructions video and you can get our Biophys Journal paper with the pictures and results on my profile.
https://www.youtube.com/watch?v=iA4Myt_PuN4
good luck!
Hello, as I saw from the video link from Margaret, I would be helpful to solve your problem.
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