I am trying to characterize normal colon stem cells from the remaining surrounding cells. Looking for techniques to etch out the differences between the two populations in the best possible manner.
the goal of the soft agar procedure is to quantify cellular anchorage-independent growth as an indicator of cellular transformation. regular adherent cells would fail to proliferate if not attached to a surface, however cancer cells wouldn't. In CFU cells have to adhere to the plate
The original CFU or CFC assays were performed in soft agar. However, whereas, soft agar is a solid matrix, methylcellulose is a viscous matrix that is water-soluble. The soft agar culture produces colonies that can be viewed as truly 3D, while methylcellulose allows the cells to go to the bottom of the plate, producing more of a 2D culture for small colonies, but a 3D culture for very large colonies. Both methods are rather insensitive, inaccurate and not particularly reliable when it comes to reproducibility. Preferred Cell Systems offers 3 different versions of the CFC methylcellulose assays, but also far more reliable, instrument-based assays, including HALO, HALO Real Time, HemoFLUOR and HemoLIGHT. Go to the website at www.preferred-cell-systems.com
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