Biomarker for predicting tumor immunotherapy response
A malignant tumor and immune checkpoint technology, applied in the field of biomarkers to predict the response of tumor immunotherapy, can solve the problem that lung cancer patients cannot benefit from it, and achieve the effect of enhancing the effect
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[0029] 1 Materials and methods
[0030] 1.1 Preparation of single cell suspension of surgically resected lung cancer samples
[0031] 1) Sample preparation
[0032] Two pairs (primary lung tumor [pT1N0M0] and matched normal lung tissue) lung samples were collected and immediately transferred for single cell isolation. Tumor tissue and normal lung tissue adjacent to the tumor were obtained during surgery at Huadong Hospital (Shanghai, China). Tumor samples were collected from three different sites in the tumor bed. The normal lung tissue was at least 5 cm away from the malignant tumor area. Tissues were rinsed with cold PBS (Cytiva, SH30256.01) to wash away blood and dead cells. Then, they were placed in 1.5 mL of chilled tissue storage solution (Miltenyi, 130-100-008) and transferred to the laboratory on ice. Mince tissue into small pieces (less than 1 mm) within 5 minutes 3), adding digestion buffer (4.7 mL DMEM (Cytiva, SH30243.01) + 325 μL enzyme mix (Miltenyi, 130-09...
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