Our colleagues are preparing for their 3D spheroid culture and they plan to use 1~1.5% agarose solution to coat the plate. When choosing the agarose for use, there are different kinds to agarose with different melting point, gelling points and EEO properties. There is no guidance about which type of agarose is suitable for plate coating. For those with a gelling point at 36 ℃, we have some concerns about melting of agarose coating at 37 ℃ in the cell culture incubator chamber. For those who have experience on this field, which type of agarose should we choose?
I have used normal PCR grade agarose and it performed well. 1% solution of agarose for PCR elecrophoresis in PBS buffer is enought to stop adhesion of cells to surface. 1g 0g agarose in 99mL of PBS buffer, autoclave and pour 200 uL of hor solution on the well of 24 well plate. Just enought to cover all surface. Better not give so much because when You seed Your cells You will dilute culture medium as it will slowly mix with PBS in gel.
Thank you, Łukasz Sieroń ! That is very helpful information! I just found a paper: 10.1038/nprot.2008.226. It recommends A9539 agarose. Other parts are the same as you mentioned. I think most agarose types (usually gelling point at 36 deg C, mp. at ~65 deg C) should work, since they will not melt upon the gel formation easily.
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