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Reducing cell count

Reducing cell count - Molecular Biology Techniques

Reducing cell count - Molecular Biology Forum. Includes forums for common molecular biology techniques.


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  #1  
Old 12-04-2012, 06:45 PM
Pipette Filler
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Default Reducing cell count



Hi,

I am currently trying to culture suspension monomac-6 cells (monocytic). They appear to be increasing in number under the microscope however when counting them they continually seem to drastically reduce. As the coutler counter bases counts on particles within solution I am not sure whether it will register dead cells in the counts. If it does count dead cells then I am perplexed as to where these cells are disappearing to. I have checked the centrifuge speed and I dont think its that.
If anyone has had any experience culturing these cells or have any idea where I am going wrong it would be much appreciated!

Andrew
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Old 12-05-2012, 03:32 AM
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Default Re: Reducing cell count

Hi,

monomac-6 cells are seeded at a density of 2 x 10 5 /ml in 2 ml volumes in 24-well-plates and they are cultured at 37C in a humidified 5% CO 2 atmosphere for 3-4 days before being used for experiments or before being pooled, counted and reseeded again (for washing spin at 400 g for 7 min). Doubling time is approx. 2 days. The culture medium is RPMI 1640 with 10-20 % fetal calf serum, fortified with several additives.

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Old 12-05-2012, 10:12 AM
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Default Re: Reducing cell count

Thanks,

Ill give that a try and let you know how I get on.
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