Journal: Stem cell research & therapy

Article Title: Regulation of self-renewal and senescence in primitive mesenchymal stem cells by Wnt and TGFβ signaling.

doi: 10.1186/s13287-023-03533-y

Figure Lengend Snippet: Fig. 2 Proliferation and colony-forming efficiency assays of LP and HP pMSCs grown in FM and XM. a Doubling time of pMSCs b, c Crystal violet stained colonies of pMSCs and phase contrast showing the morphology of a single colony of pMSCs performed in triplicate. Scale bars represent 100 μm (magnification: 4×). d Percentage of colony formation of LP and HP pMSCs performed in triplicate. E Proliferation of pMSCs as determined using BrdU proliferation kit performed in triplicate. f Relative telomerase activity in various cell types as determined using the qRT-PCR-based TRAP assay performed in triplicate. All values are reported as telomerase activity relative to HEK cells. Growth of pMSCs in FM yielded cells with significantly reduced doubling time and CFE, less compact colonies, and decreased proliferation rate and relative telomerase activity. g Light microscopy images of SA-β-gal stained pMSCs. Blue staining indicates senescent cells. Scale bars represent 100 μm (magnification: 4×). Any results showing **p ≤ 0.01 and *p ≤ 0.05 were deemed statistically significant

Article Snippet: To measure their proliferation, we followed the manufacturer’s guidelines and used the BrdU proliferation kit in triplicate (Novus Biologicals, Centennial, CO, USA).

Techniques: Staining, Activity Assay, Quantitative RT-PCR, TRAP Assay, Light Microscopy