Optimization of Culture Medium and Transfection Method for Head and Neck Squamous Cell Carcinoma Organoids  

作　　者：Zhongheng HUANG Xi YAO Qi LIU Ying XIE Zhengbo WEI 

机构地区：[1]Department of Head and Neck Surgery,Guangxi Medical University Cancer Hospital,Nanning 530021,China [2]Life Sciences Institute,Guangxi Medical University,Nanning 530021,China [3]Guangxi Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor,Nanning 530021,China

出　　处：《Medicinal Plant》2023年第3期100-104,共5页药用植物：英文版

基　　金：Supported by Natural Science Foundation of China(82160386);Guangxi Natural Science Foundation(2023GXNSFAA026189;2021GXNSFAA075042)。

摘　　要：[Objectives] To optimize the culture medium for head and neck squamous cell carcinoma patient-derived organoid and screen suitable cytokines;compare the transfection efficiency of direct transfection and short-term suspension transfection for organoid in matrigel. [Methods] Advanced DMEM/F12 medium, GlutaMax and HEPES buffer, nicotinamide, N-acetylcysteine, B27, A83-01, EGF, Y-27632 and Primocin primary cell antibiotics were prepared. On this basis, fibroblast growth factor 10(FGF10), Neuregulin 1, Noggin and R-spondin-1 were added in turn to prepare the selection medium, and the organoid diameter was used as the evaluation index to evaluate the effect of organoid medium. Using lentivirus, mCherry red fluorescent protein was transfected into HNSCC—PDO in different ways, and the transfection effect was evaluated by the fluorescence intensity of organoid sphere. [Results] Nrg1 Noggin and R-Spondin-1 promoted the growth of head and neck squamous cell carcinoma sphere(P<0.05) while FGF10 did not significantly promote the growth of head and neck squamous cell carcinoma sphere(P>0.05). Compared with direct transfection, short-term suspension transfection had higher transfection efficiency for HNSCC—PDO in matrigel. [Conclusions] R-Spondin-1 Nrg1 and Noggin may be the key cytokines in culture of HNSCC—PDO whereas FGF10 played an insignificant role in this study. Short-term suspension transfection could improve the transfection efficiency of lentivirus to HNSCC—PDO.

关 键 词：Head and neck squamous cell carcinoma Organoid culture Organoid transfection 

分 类 号：R739.91[医药卫生—肿瘤]