Journal article Open Access

Optimisation of seeding of wild-type and FOP fibroblasts for an alkaline phosphatase assay using calcein AM staining

Brown, Elizabeth; Bullock, Alex

C2C12 cells did not produce a large fold change in ALP expression following ligand administration (see previous post), therefore we will try to use FOP fibroblasts instead as these have a greater signalling response to BMP ligands. In order to complete this assay I must work out how many cells to seed in order to get a confluent well after 7 days of treatment (the expected period of time required for later compound assays). As the cells were very pale I employed a Calcein AM/Hoechst 3342 stain to mark viable cells.

Funding Acknowledgment: The SGC is a registered charity (number 1097737) that receives funds from AbbVie, Bayer Pharma AG, Boehringer Ingelheim, Canada Foundation for Innovation, Eshelman Institute for Innovation, Genome Canada through Ontario Genomics Institute [OGI-055], Innovative Medicines Initiative (EU/EFPIA) [ULTRA-DD grant no. 115766], Janssen, Merck KGaA, Darmstadt, Germany, MSD, Novartis Pharma AG, Ontario Ministry of Research, Innovation and Science (MRIS), Pfizer, São Paulo Research Foundation-FAPESP, Takeda, and Wellcome.
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