I saw some protocol that used diluted Matrigel (1 to 1 Matrigel with DMEM/F12.) for epithelial cell cultrue. I know that diluted matrigels are used for plate coating, however is there someone who used this method for epithelial cell culture such as Organoid culture?
Hope you share some experience with me!
Hi
I used diluted matrigel for epithelial cells clonogenic assays and yes 1:1 is fine
best.
Celimene
Go through the technical datasheet of Matrigel. Low concentration matrigel conc is roughly 8-9mg/ml. High concentration matrigel 20-22mg/ml. If you want to have a thin coating of matrigel on the well of your culture plate you can use diluted matrigel 1mg/ml. Remember dilute matrigel in RPMI1640 media maintained at temperature less than 4 degree C. Add the diluted matrigel to your well so that it can cover the entire surface eg. 300ul for a well of 24well, 600ul for a well of 12 well, 1ml for a well of 6 well plate. Incubate the plate at 37 deg C for at least 30 minutes. After incubation was the well with PBS kept at room temperature. Do not allow the well to dry.
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