Large scale macrophage isolation from different donors?

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sammle
sammle's picture
Large scale macrophage isolation from different donors?

Hi,
I will do a large scale macrophage isolation in the near future and would like to pool the monocytes from donors (prob.)with different blood types.
I am using the Ficoll and adherent cells protocol. Well theoritically there shouldn't be any problem but I'm worried that my isolation could still have trace of contamination by other cells that may interact.
Before doing a mistake, I would like to know if any of you have had the experience with macrophages from different blood types. Can I just mix them together?

heehawmcduff
heehawmcduff's picture
I know that cells from

I know that cells from different donors would almost certainly have differences in their response to a pro-inflammatory stimulus eg. LPS where you are measuring TNF production.  The other thing that you may have to consider is that certain people may have mature macrophages that are skewed towards either a Th1 or Th2 phenotype.  However, I guess, depending on what you are doing, if you have a large enough pool of donors then you may be able to circumvent donor-specific differences.

Kvachhani
Kvachhani's picture
hi,

hi,
just try initialy with two to three different blood group and mix it if result is possitive you can go further.
Or after separating the machrophage from different donner separately, just mix very small quantity / number of the machrophage from with different blood group and see the result which will give you idea for further stape.
regards.

sammle
sammle's picture
Hi,

Hi,
thanks for the answers..... @heehawnmcduff: well, that was actually the reason why I'm trying to pool the macrophages ;-)
I already mixed a small amount anyway, and it looked okay after 5 days. I'll see whether they will stay healthy later on.

heehawmcduff
heehawmcduff's picture
Sorry about that - I

Sorry about that - I misunderstood - I have pooled monos previously for some negative controls in expts and haven't noticed any adverse effects (using pooled autologous serum too)
Hope it works out
Best

Kvachhani
Kvachhani's picture
one of my friend done mixing

one of my friend done mixing of lymphocyte from different donner, according to him it works.
More you have tried small quantity and till five days you didnt find any problem means it will work, but watch another few days.
hope this will work, best luck.
Regards.