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[Question] how to make primary-cultured cells evenly distributed? - (Aug/05/2012 )

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Also, when you are pipetting into the chamber, are you adding a small volume of cells to a greater volume of media?

If so, instead I would suspend the cells in the total volume first- then add the whole lot to the chamber at once. This eliminates the need for mixing/shaking/rocking/whatever completely- as the cells are already evenly mixed in your suspension.


Thanks for your attached file, bob1. Since this unevenly distribution is only seen in cardiomyocyte culture but not in my other cell cultures in the same incubator, I feel it may not be due to the mechanic vibration. I attached a sketch of the uneven cell seeding I encountered, trying to show the two problems when using a 4-chamber slide, tangled cells in patches (or maybe not digested completely? How do I know the digestion is just enough?) most observed in the middle of the chamber, and cells landed heavily around the well ( Was the way I shake the chamber slide wrong? It seems hard to handle containers with small wells, such as 24 well plates.)
Leelee, I use an intelli-mixer to rotate the tissuses at the speed of 4 RPM. After filtered the cells, however, I didn’t do so many times of pipetting as you said. I’ll pay attention to this next time and see if it helps, thanks.
And thanks you all, dear forum fellows.
Attached Image

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