I am growing MEFs in a T75 flask in 15 ml high glucose and 10% FBS media each time. I started off with 2 million cells and they are reaching 70-80% confluency at the expected rate (within 2-3 days) however the cell number count (using hemocytometer) is coming out low, i.e still 2-3 million cells.
What number would you expect with MEFs specifically, general cell counts are around 8 million I think for confluent T75 flask, but I'm wondering if this is an overestimation for fibroblast cells considering their morphology.
Any ideas as to why I might be losing cells? (besides trypsin step...which is also a trouble shooting option)
Dear Selin Yildizoglu,
I am not quite sure, but it seems there was a similar question with answers. The following link.
https://www.researchgate.net/post/What_is_the_density_of_MEFs_at_100_confluency
Hi Selin. Were you able find an answer to your question?
I have similar worries. I have been trying to achieve higher numbers in a T75 80 % confluent flask, but the cells seems to die over the days and my count is either less than the seeding count or same.
Maybe you can help me with the issue based on your MEF culture experience. Thanks in advance.
Ayeshah A Rosdah at what passage were your MEFs? Were your MEFs immortalised?
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