Fig. 2: Role of calcium on MSCs viability and cell proliferation.

a MTT assays were performed to investigate cell viability is shown as bar graph (expressed as mean ± standard error) or Liner plot. MSCs cells were treated with 5 mM EDTA (for 0 mM Ca²⁺) and increasing doses of calcium for 1 and 3 days and MTT assays were performed. b Colony-forming unit (CFU) assay shows the stem cell potential after incubation with different calcium concentrations as labeled in the figure. CFU microscopic images and line-graph show the amount of stem cell potential with increasing Ca²⁺ concentrations. c Cell-cycle analysis was performed to study the different phases of the cell cycle after varying calcium treatment. The histogram and quantification of the data (shown as bar graph expressed as mean ± standard error) show the increment in the cell growth phase, i.e., S phase, was increased with increasing Ca²⁺ concentrations d BrdU assay was performed to study the cell proliferation of MSCs after treatment (1 and 3 days) with 5 mM EDTA and different doses of calcium, which is expressed as mean ± standard error from four independent experiments. e Microscopic images showing MSCs differentiation into adipocytes and osteocytes lineages in various calcium concentrations. Mineral deposition (Van Kossa staining) and lipid droplets (oil red staining) was used to evaluate osteocyte and adipocyte differentiation respectively. NS indicates data not significant and *p ≤ 0.001 (Student’s t-test).