Nanoglo substrate kit

Example 29

In the following example, NSG mice were implanted with OVCAR8-fLUC cells IP. 14-21 days after tumor implantation, 1×10⁶ human BM-MSCs engineered to express Nanoluc-EGFP were delivered IP. Mice were euthanized at 24 hours post injection of MSCs and peritoneal organs (stomach, kidney, liver, colon, spleen, pancreas, omentum/tumor, ovaries and Fallopian tubes) were imaged ex-vivo for NanoLuc signaling (NanoGlo Substrate Kit, Vendor: Promega, Catalog No.: N1110). Human MSCs were imaged by EGFP fluorescence in tumor sections collected at 24 hours as well as 22 days post injection.

As shown in FIG. 61A and FIG. 61B, human MSC NanoLuc signal was preferentially enriched in the tumor compared to the other organs in the peritoneal cavity (FIG. 61A summarized luciferase quantification; FIG. 61B representative images of luciferase signal). Additionally, persistence of MSCs was lower than 22 days, with no cells being detected at the latest time point (FIG. 61B right most panel).

Example 11

In the following example, balb/c MSCs (2×10⁶ cells) expressing fLUC were injected IP into CT-26 IP tumor-bearing mice. Mice were euthanized and tissues were collected 24 hours after injection. As shown in FIG. 37, fLUC-MSCs were significantly enriched in the tumors as detected by bioluminescence imaging (images shown in FIG. 37A, quantification of images in FIG. 37B), quantitative real time PCR (FIG. 37C), and fluorescence microscopy against firefly luciferase (FIG. 37D).

Additionally, C57Bl/6 mice were implanted with 5×10⁴ B16F10-fLUC cells IP. 7 days after tumor implantation, 1×10⁶ C57Bl/6 murine BM-MSCs engineered to express Nanoluc-EGFP were injected IP. Mice were euthanized at 24 hours post injection of MSCs and peritoneal organs (stomach, kidney, liver, colon, spleen, pancreas, omentum/tumor, ovaries and Fallopian tubes) were imaged ex-vivo for nanoluc signaling (NanoGlo Substrate Kit, Vendor: Promega, Catalog No.: N1110). As shown in FIG. 37E, murine MSC nanoluc signal was preferentially enriched in the tumor compared to the other organs in the peritoneal cavity in a B16F10 tumor model.

Example 11

In the following example, balb/c MSCs (2×10⁶ cells) expressing fLUC were injected IP into CT-26 IP tumor-bearing mice. Mice were euthanized and tissues were collected 24 hours after injection. As shown in FIG. 37, fLUC-MSCs were significantly enriched in the tumors as detected by bioluminescence imaging (images shown in FIG. 37A, quantification of images in FIG. 37B), quantitative real time PCR (FIG. 37C), and fluorescence microscopy against firefly luciferase (FIG. 37D).

Additionally, C57Bl/6 mice were implanted with 5×10⁴ B16F10-fLUC cells IP. 7 days after tumor implantation, 1×10⁶ C57Bl/6 murine BM-MSCs engineered to express Nanoluc-EGFP were injected IP. Mice were euthanized at 24 hours post injection of MSCs and peritoneal organs (stomach, kidney, liver, colon, spleen, pancreas, omentum/tumor, ovaries and Fallopian tubes) were imaged ex-vivo for nanoluc signaling (NanoGlo Substrate Kit, Vendor: Promega, Catalog No.: N1110). As shown in FIG. 37E, murine MSC nanoluc signal was preferentially enriched in the tumor compared to the other organs in the peritoneal cavity in a B16F10 tumor model.