Especially for longer-term experiments (i.e., multiple days), it is important to completely surround each well with 100 μl of water. This reduces dehydration of media in the microwells and maintains the health of the cultures. For cultures extending longer than 72 h, the cultures are replenished by removing 90 μl of media and replacing with 100 μl of media per well. This medium is slightly different than that used above to start the assay: Medium 199 containing a 1:250 dilution of RS II, ascorbic acid at 50 μg/ml, as well as 80 ng/ml of FGF-2, and 40 ng/ml each of SCF, IL-3, and SDF-1α.
Especially for longer-term experiments (i.e., multiple days), it is important to completely <br>surround each well with 100 μl of water. This reduces dehydration of media in the <br>microwells and maintains the health of the cultures. For cultures extending longer than 72 h, <br>the cultures are replenished by removing 90 μl of media and replacing with 100 μl of media <br>per well. This medium is slightly different than that used above to start the assay: Medium <br>199 containing a 1:250 dilution of RS II, ascorbic acid at 50 μg/ml, as well as 80 ng/ml of <br>FGF-2, and 40 ng/ml each of SCF, IL-3, and SDF-1α.
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