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growing macrophages - (Jan/28/2010 )

hello, i have a very basic question about macrophage growing: i know that the j774.2 doubling time is ~20 hrs...do continous cells reach a plateau in terms of growth or do they just keep growing?
Basically, 2 weeks later would I have around 50 flasks?

If anyone has any experience with the growth rate of macrophages - please share your experience! I am a newbie to cell culture. <_<

-prof. moriarty-

prof. moriarty on Jan 29 2010, 07:36 AM said:

hello, i have a very basic question about macrophage growing: i know that the j774.2 doubling time is ~20 hrs...do continous cells reach a plateau in terms of growth or do they just keep growing?
Basically, 2 weeks later would I have around 50 flasks?

If anyone has any experience with the growth rate of macrophages - please share your experience! I am a newbie to cell culture. <_<



J774 murine macrophages grow like are the easiest cells to grow. We grow them in suspension culture using Techne Stirrer bottles. Basically if you have a 500ml stirrer, you can seed that with 1ml of cell suspension at 1,000,000 cells/ml. In 3 days you should have 500ml at approximately 600,000-800,000 cells/ml. The viability should always be 99%.
We use stirrers because the cells stick to TC plastic like superglue and cannot be easilt trypsinised without increases non viability drastically.

Hope this is useful.

Rhombus

-rhombus-

Thank you for the info, Rhombus! However, I am growing them in flasks. Do you have any experience with this? How fast do they grow in flasks?

-prof. moriarty-

prof. moriarty on Jan 30 2010, 12:54 AM said:

Thank you for the info, Rhombus! However, I am growing them in flasks. Do you have any experience with this? How fast do they grow in flasks?



As stated before, if your J774 are healthy then they are very difficult to remove via trypsinisation. The trypsin stays on the cells for a long time and a loss in viability results.
The growth kinetics are similar whether they are grown in suspension or on plastic. We tried years ago to try and find a TC plastic that the cells did not adhere to as firmly...however every TC plastic was found to be the same.

Regards


Uncle Rhombus

-rhombus-