mouse macrophage cell line - (Apr/05/2010 )

Anyone know of any adherent mouse macrophage cell lines that are relatively easy to work with (in terms of passaging/maintenance)

Thanks!

RAW 247 is an adherent macrophage cell line. It is easy to work with as it can be passaged by scraping and so doesn't require trypsinisation

You could also use J774 but I believe RAWs are easier to maintain.

prof. moriarty on Apr 6 2010, 03:33 AM said:

J774 and RAW 264.7 are very similar. We have used both cell lines in our studies with Inducible Nitric Oxide Synthase (iNOS). They are as rightly stated an "adherent" cell line. HOWEVER...they both stick to TC plastic like superglue and therefore are very hard to trypsinize. Thus researchers generally scrape them as SuMi states. BUT this does reduce the viability of the cells, as scraping damages a large percentage of them.
I have stated many times on this forum about growing them in suspension using Techne Stirrer bottles......100% viability and NO TRYPSINIZATION. Both these cell lines can be grown in the same way. The initial cost of the stirrer bottles and platforms can be outweighed by the cost saving on TC plastics, trypsin and PBS "A" over time.

Hope this is useful

KIndest regards

Uncle Rhombus

P.S. One of our papers...... Moncada et al Biochem.J. (2000) 346, 407-412. Continous exposure to high concentrations of Nitric Oxide leads to a persistent inhibition of oxygen consumption by J774 cells as well as extraction of oxygen by the extracellular medium.

strange, i obtained my cells from the ECACC (through Sigma Aldrich) and in their description of the cell line they state that it is semi adherent...you can look it up on the sigma website, the cell line is j774.2 mouse macrophage.

rhombus, would you know of any effects on protein expression by growing them in spinner bottles as opposed to flasks? i am conducting a microarray of these cells when stimulated.

thanks for the reference, by the way.

prof. moriarty on Apr 9 2010, 07:15 AM said:

I used to produce Inducible Nitric Oxide Synthase (iNOS) enzyme by inducing them with Interferon Gamma and LPS. This was used for drug screening in a large pharmacuetical company. I induced the J774.1 (ATCC TIB 67) in stirrer bottles and found that I could produce more enzyme that way.....also in a very small footprint i.e. I would have had to seed 50 T175cm flasks to achive tthe same amount of enzyme, taking up a full incubator in the process.

If you are doing Gene Microarrays then my guess is that you would get slightly different array profiles. The main reason for this is that the cells are grown in suspension and are therefore exposed to "shear Stresses" that cells grown adherently are not. However I would say that the precursors of macrophages (leukocytes) travel in the blood in suspension. Differentiated macrophages normally are found in tissues engulfing foreign matter. Thus which is more relevant......suspension or non suspension? Which cellular model is correct. I would suggest looking at both.


Sorry another refernce for you from our group:-

Moncada et al Biochem&Biophys. Resarch communications 221 37-41 (1996): Visualisation of Nitric Oxide by Activated Murine Macrophages.

Kindest regards

Uncle Rhombus