Establishment and characterization of immune cells in an orthotopic cell-derived xenograft mouse model of human lung cancer using multi-color flow cytometry

doi:10.3969/j.issn.1006-7795.2023.01.015

Journal of Capital Medical University ›› 2023, Vol. 44 ›› Issue (1): 99-106.doi: 10.3969/j.issn.1006-7795.2023.01.015

• Basic Research • Previous Articles Next Articles

Establishment and characterization of immune cells in an orthotopic cell-derived xenograft mouse model of human lung cancer using multi-color flow cytometry

Yang Min¹, Yan Xiang^2*, Liu Xijuan³, An Guo⁴, Ding Huirong³, Cai Ying⁵, Ge Huizheng⁵

1. Department of Gerontology, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China;
2. Department of Medical Oncology, The Fifth Medical Center of PLA General Hospital,Beijing 100039, China;
3. Department of Central Laboratory, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Peking University Cancer Hospital & Institute, Beijing 100142, China;
4. Department of Central Laboratory, Department of Laboratory Animal Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Peking University Cancer Hospital & Institute,Beijing 100142, China;
5. Center of Excellence(Beijing), Becton Dickinson Medical Devices (Shanghai) Co Ltd., Beijing 100176,China

Received:2022-10-18 Online:2023-02-21 Published:2023-01-13
Contact: *E-mail:yxiang301@sina.com
Supported by:
National Natural Science Foundation of China (81902910).

Abstract

Abstract: Objective To establish a method to classify T cells, macrophages, and natural killer (NK) cells with detail in tumor microenvironment are needed to address the factors affecting tumor cell progression. Methods The combination of 11-color multiple fluorescence labeling markers for epithelial cells, T cells, macrophages, and NK cells was established to identify blood samples from immunized mice. Luciferase labeling PC-9 cells were further inoculated in the lung of nude mice. More real M1 and M2 macrophages and NK cells of different maturity levels were identified with penal analysis. Results This strategy can maximize the fluorescence effect of various markers and reduce the color cross-talking. Flow cytometry analysis of mouse blood samples allows accurate classification and typing of lymphocytes, macrophages, and NK cells. In orthotopic cell derived xenograft(CDX) samples, tumor cells, various immune cells, and cell subtypes can be distinguished. After the oncolytic virus treatment, the immune cell types were found significantly different. Conclusion In the orthotopic CDX model, the 11-color flow cytometry(FCM) scheme can accurately classify various immune cells in the tumor microenvironment, showing potential in the study of the interaction between different immune cells and tumor cells, and further apply to tumor microenvironment studies.

Key words: multi-color flow cytometry, lung cancer,orthotopic transplantation, immune microenvironment, immune cells, oncolytic virus

CLC Number:

Yang Min, Yan Xiang, Liu Xijuan, An Guo, Ding Huirong, Cai Ying, Ge Huizheng. Establishment and characterization of immune cells in an orthotopic cell-derived xenograft mouse model of human lung cancer using multi-color flow cytometry[J]. Journal of Capital Medical University, 2023, 44(1): 99-106.

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Establishment and characterization of immune cells in an orthotopic cell-derived xenograft mouse model of human lung cancer using multi-color flow cytometry

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