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Protocol for high-throughput droplet-engineered TIL-retaining tumor organoids in triple-negative breast cancer

Xiaolong Zhou, Yu Zhu, Wanlong Wang, Yongde Cai, Zhiyuan Liu, Yingshi Su, Shaohua Ma

Journal:STAR Protocols

IF:1.4

DOI:10.1016/j.xpro.2026.104706

PMID:42470642

Published:2026-07-17

research field:肿瘤学分子生物学心血管生物学遗传学与基因组学

Abstract

Summary Here, we present a protocol for generating high-throughput, droplet-engineered tumor organoids from triple-negative breast cancer (TNBC) tissues using the OrgFab® micro-sphere printing system. We describe tumor dissociation, bioink preparation, and automated organoid fabrication. We further detail rho-associated coiled-coil kinase (ROCK) pathway modulation to preserve tumor-infiltrating lymphocytes (TILs) and enhance T cell function, as well as flow cytometry-based immune characterization ( Figure 1 ). This protocol enables scalable production of immune-competent tumor organoids for immunological analysis and drug-screening applications. For complete details on the use and execution of this protocol, please refer to Zhou et al. 1

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