Research Paper Volume 11, Issue 20 pp 9167—9187

Bone marrow-derived mesenchymal stem cells in three-dimensional co-culture attenuate degeneration of nucleus pulposus cells

Figure 1. Mesenchymal stem cells activated in IDD, and cocultured BMSCs can alleviated the nucleus pulposus cells senescent rate in vitro. (A) Immunohistochemistry staining of CD44 in intervertebral disc degeneration of Pfirrman grade III(G III) and grade IV(G IV). n=5, Scale bar, 50um. (B) CD44 positive cell rate between G III and G IV were determined by using Image J software. (C) SA-β-Gal staining of primary NP cells of IDD Pfirrman grade IV and Pfirrman grade III. n=5, Scale bar, 50um. (D) SA-β-Gal positive cell rate between G III and G IV were determined by using Image J software. (E) 3D coculture models of were established as schematic diagram described. SA-β-Gal staining of senescent NP cells after 2 days coculture of normal NP cells +blank calcium alginate gel balls(CTL), senescent NP cells+ blank calcium alginate gel balls(Sene), senescent NP cells +calcium alginate gel balls with BMSCs(Sene +BMSCs), senescent NP cells +calcium alginate gel balls with normal NP cells(Sene +NPCs), n=3, Scale bar, 50um. (F) Co-cultivation pattern design and experimental groups. (G) SA-β-Gal positive cell rate between CTL, Sene, Sene +BMSCs and Sene +NPCs groups were determined by using Image J software. (H) Experimental flow diagram Values represent means±S.D. Significant differences between different groups are indicated as *P < 0.05, **P < 0.01, ***P < 0.001. PF: per field.