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. Author manuscript; available in PMC: 2019 Apr 22.
Published in final edited form as: Nat Biomed Eng. 2018 Oct 22;2(12):915–929. doi: 10.1038/s41551-018-0307-x

Fig. 4. Instigation of humanized pre-metastatic niches with hPBMCs and long-term monitoring of DTC niche evolution via serial transplantation.

Fig. 4.

a, Experimental schematic describing the establishment of early stage humanized DTC niches and subsequent instigation by systemic introduction of hPBMCs. b, Representative IHS of humanized early DTC niche from 3 independent experiments including hCytokeratin (tumor), hCD45 (immune), hVimentin (stroma) and mCD31 (vessel). Arrows indicate human immune (white) and tumor (green) cells. c, Representative ex vivo BLI of explanted scaffolds from primary mice and quantified bioluminescent signal with and without hPBMCs. The dotted line represents a threshold used to define BLI+ and BLI-. Data are mean ± s.d. Not significant, P > 0.05 via two-sided student t-test. d, Experimental schematic describing a strategy to monitor the long-term evolution of humanized DTC niches via serial transplantation of intact early metastatic niches to naïve secondary syngeneic mice. e, Representative whole body BLI of primary and secondary mice at different time points from 2 independent experiments. Gross image of explanted scaffolds from secondary mice showing an overt metastasis. f, Quantitative analysis of human DTC growth in scaffolds after transplantation into secondary mice via weekly BLI. Scaffolds were classified as BLI-, BLI+, or overt based on their bioluminescent intensity. Presented N values are representative of independent scaffolds.