We are trying to grow the U251 and U87 cells in Neurosphers. At first when I cultured the cells in DMEM high glucose with 1% PEST, 1% L-Glut and 1% NEAA with 10% regular FBS (non exosome depleted), they grew beautifully into neurospheres on agar coated plates, but when I changed the medium to exosomes depleted FBS (commercially available from GIBCO), the cells started to form monolayers on Agar. I used a mild dissociation Accutase for passaging these cells. In addition, the monolayer culture in both regular FBS and exosome deplted FBS did not alter the cellular characteristics. We are still wondering what could be the possible reason for these cells not to form neurospheres on agar plates with exosomes depleted medium.
PS: The regular FBS that was used primarily was from SIGMA and the exosome depleted FBS was from GIBCO.
Answers and comments would be appreciated. Thanks
Taral
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