Figure 1
From: Role of IGF1R+ MSCs in modulating neuroplasticity via CXCR4 cross-interaction
Isolation and clonal expansion of IGF1R-expressed hDSCs.
(a) In the representative figure of cytokines array (square P = positive control, square N = negative control), five different cytokines such as EGF (square #1), Angiogenin (#2), MIP-1δ (#3), RANTES (#4), and PDGF-BB (arrow) were significantly up-regulated in hUCS compared to FCS. The level of IGF1 (arrow head) was almost the same between hUCS and FCS. (b) In ELISA, the concentration of serum PDGF-BB showed significant increase in the hUCS compared to that of FCS, but not found in serum IGF1. (c) Fibroblast-like cell morphology (arrow) was found in hDSCs from dental pulp explant (arrow head, left panel). Surface marker of IGF1R and other MSCs’ markers was not only found in hDSCs, but in human fibroblast (positive control), ADMSCs, UMSCs and BMSCs. In flowcytometric analysis, hDSCs carried the surface markers of MSCs and pluripotent markers of Oct-4, Sox-2, Nanog and SSEA-4. (d) hDSCs were cultured in 2% hUCS or 10% FCS. In representative graph of the expansion potential profile, U-IGF1R+ hDSCs revealed more fast growth rate than that of U-IGF1R‒ hDSCs and U-IGF1R+ hDSCs+PPP. hUCS-cultured hDSCs revealed rapider division than FCS-cultured hDSCs. In addition, higher cellular proliferation rate was abolished in both hUCS-cultured hDSCs (U-hDSCs) and FCS-cultured hDSCs (F-hDSCs) infected with LV-IGF1R-shRNA. (e) Western blot exhibited significant upregulation of Bmi1 and cdk4, as well as downregulation of p21 in U-IGF1R+ hDSCs compared to that of U-IGF1R‒ hDSCs, but no difference in cyclinD1, JunB and PTEN. (f) In BrdU ELISA, U-IGF1R+ hDSCs showed significant increaser BrdU uptake compared to that of U-IGF1R‒ hDSCs and U-IGF1R+ hDSCs+PPP. More, hUCS-cultured hDSCs showed significantly increased BrdU incorporation compared to that of FCS-cultured hDSCs (SDF-1α as positive control), but not in LV-IGF1R-sh-hUCS-hDSCs and LV-IGF1R-sh-FCS-hDSCs. (g) Co-expression of IGF1R with self-renewal markers Oct-4, Sox-2, Nanog, and SSEA4 was found on hDSCs by flowcytometry. Furthermore, qRT-PCR analysis of relative expression of Oct-4, Sox-2, Nanog and SSEA4 significantly increased in p5 IGF1R+ hDSCs compared to that in IGF1R– hDSCs. Data are expressed as mean ± SEM. *P < 0.05 and **P < 0.01 vs. control, Bar = 40 μm.
