2.5. Measuring total collagen synthesis
The total collagen synthesis exhibited by the cells was measured using a Sircol™
soluble collagen assay kit, which was modified based on a previous study
(Chiang et al., 2013a,b). In brief, the cell culture medium was collected, mixed with
an isolation and concentration reagent, and incubated. The Sircol™ dye was added
to the sample and the mixture was incubated. After centrifugation, an ice-cold acidsalt
washing reagent was added to the precipitate and the mixture was recentrifuged.
The precipitate was dissolved with an alkali reagent and the absorbance
was determined at 555 nm by using an ELISA reader (Tecan, Grödig, Austria).