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In the absence of the strong growth factor stimulation found in standard proliferation media, fibroblasts growing in CnT-PR-FH show heighted responsiveness to experimental stimulii (for example Vitamin C to stimulate collagen secretion, as shown at left).
Fibroblast responsiveness to vitamin C treatment (100 ug/mL) is 5x higher than in the standard CnT-PR-F proliferation medium. Confluent dermal fibroblasts were switched to the respective media at confluence, and grown for a further 5 days. Total collagen was then lysed in the well (acetic acid), quantified using sirius red, and normalised to gDNA to correct for any differences in cell number.
It is strongly recommended to evaluate total collagen using in-well lysis of all the cells and deposited ECM (as opposed to evaluation of only the supernatant), and to normalise for cell number using gDNA.
A complete protocol is available for download.
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