Research Paper Volume 10, Issue 1 pp 131—143

Rapamycin prevents the intervertebral disc degeneration via inhibiting differentiation and senescence of annulus fibrosus cells

Figure 1. Proliferation of rabbit AFSCs cultured in growth medium with five different conditions for 6 days. (A, F) Growth medium only (Control); (B, G) bleomycin 10 μg/ml (B-10); (C, H) bleomycin 50 μg/ml (B-50); (D, I) Bleomycin 50 μg/ml and rapamycin 25 nM (B-50 + R-25); (E, J) Bleomycin 50 μg/ml and rapamycin 50 nM (B-50 + R-50); (L) Population doubling time (PDT) of AFSCs grown in five different conditions for 6 days. The images of F, G, H, I, J were enlarged areas of the boxes in images of A, B, C, D, E. Bleomycin treatment changed the morphology of AFSCs from cobblestone-like cells (green arrows in A, F) to pancake-like cells (red arrows in B, C, G, H), and also decreased proliferation of AFSCs as evidenced by population doubling time (L). However, the addition of rapamycin into bleomycin-containing medium decreased pancake-like cell numbers and increased spindle-like cell numbers (yellow arrows in D, E, I, J). White bars: 100 μm, yellow bars: 50 μm. *p<0.05 compared to control.