Chapter 7

Primary Cell Culture and T-cell Cloning - Fundamental of Adoptive T-cell Immunotherapy

Biaoru Li, Supriya Perabekam, Alan Larson and Hong-Liang Hu

Abstract

Tumor-associated antigen (TAA) or tumor-specific antigen (TSA) is essential for the target of tumor-specific T-cells such as tumor-infiltrating T cells (TIL), specific T-cells, TCR T-cells and CAR-T-cells for adoptive T-cell immunotherapy. The tumor cells often accumulate hundreds of mutations and harbor several immunogenic neoantigens, and thus, the repertoire of mutation or neoantigen from patient tumor cells might need the screen to uncover for engineering these T-cells. To understand the Tcell screening and determining tumor antigen-based on primary tumor cells from an individual patient, this chapter, we focus on streamlining the process of ex vivo T-cell culture and primary tumor cell culture, T-cell cloning for tumor neoantigen-specific T cells, allowing the patient to the benefit of downstream T-cell targets. Because T-cell engineering cultures are very important methods for TIL, TCR and CAR T-cells, moreover, because using primary tumor cells isolation and cultures is very important for screening and identifying tumor antigen of patients, we first introduce primary cell culture techniques, including those developed from two-dimensional (2-D) tumor cell cultures, three-dimensional (3-D) tumor cell culture and multiple dimensional tumor cell culture (4-D cultures). These methodologies are increasingly supporting clinical oncologists to apply to tumor therapeutic agents and Ag targets for patients in the clinical laboratory. Besides, we also conclude some growth factors for T-cell cloning cultures. The chapter aims to present a foundation to adoptive T cell immunotherapy of clinical patients.

Total Pages: 97-117 (21)

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