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This punch plating method efficiently amplifies the initial culture, so that a single mouse esophagus could potentially be expanded 3.7 10 6 -fold in 100 days (Methods, Extended Data Fig

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With just three to four samples per cohort, the spread within each cohort in the PCA score plot is notably small indicating high overall reproducibility with good, unsupervised inter-cohort and phenotype discrimination

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Rotate injection sites to prevent tissue irritation from repeated injections in the same spot