As stated,the doubling time for my sf9 cells were increased:the cells were seeded at the density of 106/ml and three days later,it only reached 2*106/ml with higher than 90% viability.The cells were newly thawed and subcultured for 4-5 passages.The conditions,including temperature(27oC) and shaking speed(120rpm)followed exactly my previous experiment,which gave good results.The same frozen stock were thawed before and they grown very well. Can anyone have any ideas?
I had the same problem with my Hi5 insect cells the past few months. The maximum density it reached was only 5-6 million/mL after three days. So I asked for another fresh flask of Hi5 cells from the company, but they still can only grow to 7million/mL after three days. It's supposed to go up to 8 million/mL. One possibility could be the stocks are old. The other one could be the media is slightly different from before.
Did you ever encounter the problem that sometimes you got good yield from Sf9, but sometimes you got nothing. I've been tortured for months by the protein expression in insect cells....
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