aspects of RAW264.7 cells - (Nov/05/2010 )

Hi all,

I am working with macrophages RAW264.7 in culture. I have them growhing in T75 Flask and I worried the aspect they have...Most of them are bright and have round shape with pseudopods. But some of them are bigger and dark, and have more differentiated shape...It is normal in that cell line or not??

thanks for all.

Alvaro.

Are the bigger cells individual or cluster cells? And is your medium made with endotxin free water?

alvaro on Fri Nov 5 10:54:27 2010 said:

Dear Alvaro

It is perfectly normal....RAW264.7 have 2 morphologies:-
Round and golden (adherent)
Flat and Grey/Dark colour (adherent).

The viability of both morphologies is the same.

If you however grow them as a suspension culture i.e. Techne Biological stirrers...then the morphology is uniform (round and golden).


Hope this puts your mind at rest.

Kindest regards

Uncle Rhombus.

ok thanks, my cells are exactly as you describe...round and golden and flat and grey color in the other side. But if they are the same cell line, why grow differently??

ouhscr2 on Fri Nov 5 16:17:23 2010 said:

thanks for your answer. The bigger cells are individual and the water is endotoxin free...but anyway, I am more relaxed now with the answer of Rhombus

alvaro on Mon Nov 8 14:33:11 2010 said:

Hi Alvaro,

Yes they should be all the same but there are sub populations in all cell systems. That is why it is essential that you grow them in exactly the same way every time i.e.
Same cell density and Split ratio's
Same media/pH conditions
Same FBS/FCS
Humidified incubator conditions
5% CO2 ....regularly monitored

If any of the above/or in combination vary for any length of time then you put "selection" pressure upon them and can select for small sub populations of cells....for example if you have acid pH conditions, then the cells that survive are cells predisposed for acid conditions...they might vary in many other ways from so called "normal cells". Our group have seen this experimentally when using J774 cells (published data).


Hope this is again useful......remember that these cells are only a model and the model is no where near perfect.

Uncle Rhombus.