Generation of Murine Cancer Cell Lines

Current protocols Pub Date : 2024-10-21 DOI:10.1002/cpz1.70037

Cheryl Zhang, Anthony Lin, Xiang Chen, Yi-Chieh Nancy Du

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Abstract

Cancer cell lines are important tools to investigate the biology of cancer and test hypotheses to improve cancer treatments. A major challenge in establishing epithelial cancer cell lines is the removal of cancer-associated fibroblasts (CAFs). CAFs are abundant within the tumor microenvironment. CAFs generally proliferate faster than epithelial cancer cells in culture. CAFs can be mistakenly identified as cancer cells, especially when cancer cells display spindle-shaped morphology. Among all cancer types, pancreatic ductal adenocarcinoma (PDAC) is characterized by an abundant desmoplastic stroma. Here, we describe protocols for establishing epithelial cancer cell lines from mouse models of PDAC and verifying that they are not CAFs. The approach is cost-effective and can be used for other types of cancer. If needed, CAF cell lines can also be established and preserved using this protocol. © 2024 Wiley Periodicals LLC.

Basic Protocol 1: Isolation of cells from tumors

Basic Protocol 2: Isolation and cryopreservation of cancer cell clones

Basic Protocol 3: Assessment of the identity of cancer cell lines and CAFs by western blotting

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生成小鼠癌症细胞系。

癌症细胞系是研究癌症生物学和验证改善癌症治疗假说的重要工具。建立上皮癌细胞系的一大挑战是去除癌症相关成纤维细胞（CAFs）。CAFs 在肿瘤微环境中含量丰富。在培养过程中，CAFs 的增殖速度通常快于上皮癌细胞。CAFs 可能会被误认为是癌细胞，尤其是当癌细胞呈现纺锤形形态时。在所有癌症类型中，胰腺导管腺癌（PDAC）的特点是具有丰富的脱鳞基质。在此，我们介绍了从 PDAC 小鼠模型中建立上皮癌细胞系并验证它们不是 CAF 的方法。这种方法具有成本效益，可用于其他类型的癌症。如有需要，也可使用该方案建立和保存 CAF 细胞系。© 2024 Wiley Periodicals LLC.基本方案 1：从肿瘤中分离细胞基本方案 2：分离和冷冻保存癌细胞克隆基本方案 3：通过 Western 印迹法评估癌细胞系和 CAF 的特性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Current protocols

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