5 replies to this topic

[pheobeyue]

Hi everyone, I am a new to cell culture. I am growing some MDCK cells in T-75 flasks. However, it always got round-shaped vasculation even has been growing for several days later( I think it should pass the log phase) But I always see vascuolation from the beginning until trypsinization.
I have attached the photo here. Could anyone help me on this problem? Is that normal? What's going on for the cells?

Thanks  a  lot!

Attached Files

•  Captured_4x_Lens_MDCK_flask2.jpg   603.97K   37 downloads
•  Captured_4x_Lens_MDCK_flask3.jpg   599.49K   36 downloads

[bob1]

The cells are over confluent and are senescent and dying.  Split your cells more frequently or seed less densely.

[pheobeyue]

bob1, on Feb 22 2010, 04:37 PM, said:

The cells are over confluent and are senescent and dying.  Split your cells more frequently or seed less densely.

Over confluent? I still saw a lot of spaced among these cells.Also, my cells grow very slowly, so I left it for like 5 days to grow.
Actually I only seed like 1X10^4 in the T-75 flask

[bob1]

pheobeyue, on Feb 26 2010, 12:07 PM, said:

bob1, on Feb 22 2010, 04:37 PM, said:

The cells are over confluent and are senescent and dying.  Split your cells more frequently or seed less densely.

Over confluent? I still saw a lot of spaced among these cells.Also, my cells grow very slowly, so I left it for like 5 days to grow.
Actually I only seed like 1X10^4 in the T-75 flask

Not in the pictures you posted, in those the cells are 100% confluent.  I haven't used MDCK but it looks like they grow pretty fast if you are only seeding 10,000 cells per T-75 and they are confluent 5 days later.  Are you sure your cell counts and calculations are correct?  If they are your cells are doubling approximately every 15 hours or so, which is very quick.

[three]

Your cells do not look healthy. I regularly culture MDCK and they do grow fast but your cells are not yet confluent. Are you sure you are using the correct media? How long do you trypsinize your cells and what concentration of trypsin do you use?

[Eric]

three, on Mar 1 2010, 09:04 AM, said:

Your cells do not look healthy. I regularly culture MDCK and they do grow fast but your cells are not yet confluent. Are you sure you are using the correct media? How long do you trypsinize your cells and what concentration of trypsin do you use?

Hi body, which strain of MDCK are you using? Do you have MDCK-c11?