Isolation of Alveolar Macrophages

Charles Pegram cnpegram at acpub.duke.edu
Tue Jun 25 12:32:22 EST 1996


James Young wrote:
> 
> Hello there....I'm a research assistant at Bradley University and I am
> working with Alveolar Macrophages.  I was wondering if there was a more
> efficient way to isolate them in large numbers.  What we are doing now is
> basically a homogenization.  Is there a better way?????
> 
> Thanks in advance
> James Young
> jyoung at camelot.bradley.edu
> .

James,
  
   You don't mention what species your are trying to isolate Alveolar 
Macrophaes from but since you are preparing a homogenate I assume they 
are not human.  What worked for us back in the late 1960's and early 
1970's in rats, mice, and guinea pigs is:  Dissect out lungs with trachea 
stilled attached.  Place a canual or blunt needle into the trachea and 
tie it in place with sutures.  Attach a syringe filled with your buffer 
or media of choice and inflate lungs with it.  Do not inflate to the 
point the lung membranes rupture.  Remove syringe and allow fluid to 
drain out into sterile tube or petri dish.  repeat this procedure 4-5 
times and you will recover most of the macrophages.

Charles Pegram
cnpegram at acpub.duke.edu



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