Department of Medicine, McMaster University, Hamilton, ON L8N 3Z5, Canada.
McMaster Immunology Research Centre, McMaster University, Hamilton, ON L8S 4K1, Canada.
STAR Protoc. 2021 Nov 17;2(4):100956. doi: 10.1016/j.xpro.2021.100956. eCollection 2021 Dec 17.
Although natural killer (NK) cells have become a promising immune effector cell for chimeric antigen receptor (CAR)-based therapy, generating human CAR-NK cells with high transgene efficiency has been challenging. In this protocol, we describe how to generate CAR-NK cells with transduction efficiencies >15% from healthy donor expanded NK cells using third generation lentiviral vectors (LVs). We also show how to assess CAR-NK cell anti-tumor function using a flow cytometry-based killing assay. For complete details on the use and execution of this protocol, please refer to Portillo et al. (2021).
尽管自然杀伤 (NK) 细胞已成为嵌合抗原受体 (CAR) 为基础的治疗中很有前途的免疫效应细胞，但高效地产生具有高转导效率的人源 CAR-NK 细胞一直具有挑战性。在本方案中，我们描述了如何使用第三代慢病毒载体 (LV) 从健康供体扩增的 NK 细胞中产生转导效率>15%的 CAR-NK 细胞。我们还展示了如何使用基于流式细胞术的杀伤测定法评估 CAR-NK 细胞的抗肿瘤功能。有关该方案使用和执行的完整详细信息，请参阅 Portillo 等人。（2021 年）。
