I plate 105/cm2 CaCo-2 cells on 6/12/24 well plates. After they grown 5 days, most cells on the well begin detached from plate, but other small cells (shape looks like rounded and attached on plate) come begin to grow under or near the detached cells. This happens even coated plates. I have a concern about cells is that when I split cells, next day on the plate large and small cells (very high number) are seen on the plate.
change medium I set very low power for pumping
every two days change fresh medium
wash cells with PBS or medium
pen/strep with anti mycotic are added medium
Any suggestion!
Similarly we have same issue recently.
Therefore, we got a new cell line from ATCC again.
Do you think mycoplasma issue? or how do new cells grow as postconflunet? Any detachment or other growth/differentiation issue? Could you provide me ATCC lot number of CaCo-2 cell?
I will also order new one, but I can not learn lot number until I order in here. I can request same lot number from Attc if you test cells.
Hello
See the links here hope help you , and you can contact with ATCC about this issue
Good Luck
http://link.springer.com/chapter/10.1007%2F978-3-319-16104-4_10
https://www.researchgate.net/post/We_are_having_problems_growing_caco-2_cells_they_grow_very_slowly_Can_anyone_give_us_tips_on_how_to_make_them_grow_faster
http://www.biocompare.com/6-Tissue-Culture-Forums/1011-Caco-2-cell-culture/?ftpid_6737=4
What's the doubling time of Caco 2 cell line ? And what percentage of FBS you are using ?
Would you please attach a photo for these small cells with your question?
It would help.
This is my question:
I plate 105/cm2 CaCo-2 cells on 6/12/24 well plates. After they grown 5 days, most cells on the well begin detached from plate, but other small cells (shape looks like rounded and attached on plate) come begin to grow under or near the detached cells. This happens even coated plates. I have a concern about cells is that when I split cells, next day on the plate large and small cells (very high number) are seen on the plate.
change medium I set very low power for pumping
every two days change fresh medium
wash cells with PBS or medium
pen/strep with anti mycotic are added medium
Any suggestion!
Those are the pictures from my cells.
Dear Sukra,
Has your problem been solved? If so how did you solve this problem? I am currently encountering the same problem in regular 12- and 24-well plates. Personally I think it has to do with the formation of Caco-2 domes which eventually leads to the detachment of cells.
Thijs
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