Porcine alveolar macrophages culture - (Feb/05/2007 )

Any one can provide me a protocol for culture and maintainance of "Porcine alveolar macrophages".
Is any special culture plates Or medium required?
I am trying to culture(macrophages) in regular tissue culture plates and using RPMI medium, but I am finding less than 50% cells adherence and cells start dying after 24 hrs.
Please give me suggestions.
Thanks

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what else do you use in your medium?

is there the possibility of contamination, are the cells activated at all?

i have grown ovine alveolars in rpmi supplemented with pen/strep, 5%fcs

hope this helps

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Thanks for reply. I am using RPMI suplemented with pen/strep, 50ug gentamycin, 10%FBS. It has little chance of contamination because these are free alveolar macropgages were collected by lung lavage ( lung infiltrating by PBS).

Another thing i preserved these macrophages in Liquid Nitrogen. Or in each experiment it should use fresh.

I am using corning plates for culture.

Please let me know for changes.
Thanks

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