We shifter to new lab 8 months back. Since then all the cell lines which includes CEM, THP1 and 293T grow only and only when we seed then at high density like 6X10 to5.
This is a huge problem as for making knockdown using sh RNA we need to tale cells in the range of 10to4.
I I have not been able to carry out any experiments at all, where low cell numbers are required.
Please seggest.
Dear Namita,
I think if the cell numbers are too low they will not grow well, because they secrete growth factors and will look for cell to cell contact. If the seeding density is to low there may result in too low of concentration of growth factors to sustain a viable culture. They will get other things from the complete medium, but also will need each other to thrive.
I have not tried this but, if you was to replace/mix medium of low seeded culture with that of a well growing but sub-confluent culture, you may be able to get viable low seeding cultures.
Best,
Charles
Try using 15% FBS supplemented growth medium. Seed the cells in 15% FBS+Grwoth medium and after 24 hours you can replace the media with 10% FBS.
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