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airway primary cells stop growing at passage 3


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#1 minhduc_bio

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Posted 12 November 2012 - 03:12 AM

Dear all, I have faced a problem with my airway primary cell that: at passage 3, the cells look very healthy on the first day; The days after, the number of living and healthy cells seems to be unchanged while the number of dead cells increases (cells detach; debris appears). I kept changing medium to wash the dead cells out, but after 1 days, dead cells start appearing and increasing. The strange thing is that the healthy cells are still there but do not increase. This is the first time I have worked with primary cells so I am very confused with this problem.
Could somebody had the same problem and experience before?

#2 bob1

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Posted 12 November 2012 - 11:28 AM

How confluent are the cells? It sounds like they are contact inhibited and dying because of that.

Are they really passage 3 - as in you extracted them from the epithelium and have split them 3 times? or is it 3 passages since you thawed an aliquot? If it is the last option, have a look for information on the "Hayflick limit".




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