Anchorage Independent Transformation Assay
Do No Harm
Colony counting in soft agar is the classic assay for selecting metastatic cells. It has two problems, 1) cells can be damaged by exposing them to temperatures at and above the 40 C melting temperature of agar, 2) it can take three weeks to grow colonies large enough to count. Damaged or heat stressed cells can skew your results, especially genomics, proteomics or metabolomics analyses. Colony counting is too imprecise to wait around three weeks.
Do the Opposite
poly(2,3)Glucose is a temperature responsive 3D cell culture gel. It has the same non-adherent property of agar that selects for anchorage independent cells, but it has the opposite melting behavior. Warm poly(2,3)Glucose from room temperature to 37 C and it solidifies. Briefly chill it over ice and it melts. There is no danger of harming your cells, because the temperature never gets hotter than inside the incubator.
Do Count Cells; Do Omics; Do Replate
Recovering metastases is safe and easy with poly(2,3)Glucose. Count transformed cells with a MTT assay at week 1, instead of waiting till week 3 to get colony counts. Do omics analysis on recovered cells without concern about how heat stress may affect the results. Do replate recovered cells as often as you like.
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