Research Paper Volume 13, Issue 22 pp 24542—24559

Figure 1. Proliferative capacity of placenta-derived mesenchymal stem cells from different maternal age group. (A) Cell growth curve showing 5 age groups. Cells were seeded at 1x10⁴ per cm² in 24-wells plates and counted every 48 hours for 10 days. (B) Cumulative Doubling Population curve showing 5 age groups. Cells were seeded at 1x10⁴ per cm² in 24-wells plates and subculture every 4 to 5 days. Population Doublings (PD) was calculated using the formula ΔPD= log(total cell number/initial cell number)log2. Each dot represents one passage. Bar graph represents the cumulative population doubling among the 5 age groups at day 46. (C) MTT cell proliferation assay performed using Thiazolyl Blue Tetrazolium Bromide in a 96-well ELISA plate. (D) Graph showing Colony Forming Unit-Fibroblast (CFU-F) assay of 5 age group. 1x10³ cells were seeded in 12-wells plates and allowed to grow for 8 days, before staining with Crystal Violet. Colonies with more than 40 cells were counted. (E) Graph showing the number of cells in colonies counted for the CFU-F of 5 age groups. (F) Representative image showing cellular morphology of 5 age groups at 4 days of culture. (G) Images represent crystal violet staining in cells from each maternal age group. Group A: 18-21; Group B: 22-25; Group C: 26-30; Group D: 31-35; Group E: 36 and over. *, p<0.05; **, p<0.01; ***, p<0.0005; ****, p<0.0001. Scale bar 200 μm.