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Abstract：
A 3D bioprinted neurovascular unit (NVU) model is developed to study glioblastoma (GBM) tumor growth in a brain-like microenvironment. The NVU model includes human primary astrocytes, pericytes and brain microvascular endothelial cells, and patient-derived glioblastoma cells (JHH-520) are used for this study. Fluorescence reporters are used with confocal high content imaging to quantitate real-time microvascular network formation and tumor growth. Extensive validation of the NVU-GBM model includes immunostaining for brain relevant cellular markers and extracellular matrix components; single cell RNA sequencing (scRNAseq) to establish physiologically relevant transcriptomics changes; and secretion of NVU and GBM-relevant cytokines. The scRNAseq reveals changes in gene expression and cytokines secretion associated with wound healing/angiogenesis, including the appearance of an endothelial mesenchymal transition cell population. The NVU-GBM model is used to test 18 chemotherapeutics and anti-cancer drugs to assess the pharmacological relevance of the model and robustness for high throughput screening.
摘要：
开发了3D生物打印的神经血管单元（NVU）模型，以研究类似脑微环境中的胶质母细胞瘤（GBM）肿瘤生长。NVU模型包括人类原代星形胶质细胞，周细胞和脑微血管内皮细胞，和患者来源的胶质母细胞瘤细胞（JHH-520）用于本研究。我们使用具有共聚焦高含量成像的荧光报告分子来定量实时微血管网络形成和肿瘤生长。NVU-GBM模型的广泛验证包括对脑相关细胞标志物和细胞外基质成分的免疫染色;单细胞RNA测序以建立生理相关转录组学变化;以及NVU和GBM相关细胞因子的分泌。scRNAseq揭示了与伤口愈合/血管生成相关的基因表达和细胞因子分泌的变化。包括内皮间充质转化（EndMT）细胞群的出现。NVU-GBM模型用于测试18种化学治疗剂和抗癌药物,以评估模型的药理学相关性和高通量筛选的稳健性。本文受版权保护。保留所有权利。
