The CFU-F assay was employed as previously described17 (link) to count CTPs, whereby the bone marrow samples were added to StemMACS MSC expansion media (Miltenyi Biotec) and then cultured for 14 days. The colonies were visualized using methylene blue and counted manually. Each colony was defined as having at least 50 cells32 (link).
Protocol detail
Colony-forming unit fibroblast assay
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Corresponding Organization :
Other organizations : NIHR Leeds Musculoskeletal Biomedical Research Unit, Mansoura University, St James's University Hospital, University of Leeds, Chapel Allerton Hospital
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Variable analysis
independent variables
- Bone marrow samples
- Number of colony-forming unit fibroblasts (CFU-Fs)
- Culture time (14 days)
- Cell count threshold for defining a colony (at least 50 cells)
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