Fig. 5. EVDRL-secreting stem cells have antitumor effects against BLBC in vitro and in vivo.
(A) Left: Photomicrograph of EVDRL-secreting hMSC. Scale bar, 100 μm. Right: Concentration of EVDRL in culture media of hMSC-EVDRL quantified by enzyme-linked immunosorbent assay (ELISA) (n = 2, technical replicates). (B) Top: Photomicrographs of BMET02-FmC cocultured with hMSC-GFP/DRL/EVDRL for 72 hours. Scale bars, 100 μm. Bottom: Cell viability of BMET02-FmC after 72-hour coculture with increasing percentages of hMSC-GFP, hMSC-DRL, or hMSC-EVDRL (n = 3, technical replicates). (C) Photomicrographs of different engineered stem cells (left) (scale bars, 100 μm) and cell viability of BMET02FmC cocultured with increasing percentages (0 to 100) of the stem cells (right) (n = 3, technical replicates). (D) Left: Photomicrograph of BMET02-FmC cocultured with sECM-encapsulated hMSC-GFP, hMSC-DRL, or hMSC-EVDRL. Scale bar, 1 mm. Right: Relative number of BMET02-FmC cells 72-hour following coculture with sECM-encapsulated hMSC-GFP, hMSC-DRL, or hMSC-EVDRL (n = 3, technical replicates). (E) Left: Experimental outline for testing efficacy of sECM-encapsulated hMSC-EVDRL in BMET02-FmC–bearing mice. Right: BLI signals before and after resection (n = 20). (F) Intraoperative photographs of light and fluorescence of mice implanted sECM-hMSC into the resection cavity of BMET02-FmC tumor. Scale bars, 1 mm. (G) Representative photomicrographs of brain section from mice 2 and 4 days after resection of BMET02-FmC tumor and implantation of sECM-hMSC. Scale bars, 100 μm. (H) Estimate of relative tumor volume after resection in treatment groups based on Fluc signal intensity of BMET02-FmC (hMSC-GFP, n = 6; hMSC-DRL, n = 7; hMSC-EVDRL, n = 7). (I) Kaplan-Meier survival curves of the mice with median survival (days) indicated in the legend. (J) Immunohistochemistry of cleaved caspase-3 of brain sections from treated and control mice. Scale bars, 100 μm. Photo credit: Yohei Kitamura, Brigham and Women’s Hospital