使用聚电解质多层模板对星形胶质细胞和神经胶质瘤细胞进行直接接触共培养。

Direct-Contact Co-culture of Astrocytes and Glioblastoma Cells Patterned using Polyelectrolyte Multilayer Templates.

机构信息

Complex Biosystems Graduate Program, University of Nebraska, Lincoln; Department of Chemical and Biomolecular Engineering, University of Nebraska, Lincoln.

Complex Biosystems Graduate Program, University of Nebraska, Lincoln; Department of Chemical and Biomolecular Engineering, University of Nebraska, Lincoln; Fred & Pamela Buffett Cancer Center, University of Nebraska Medical Center; Nebraska Center for Integrated Biomolecular Communication, University of Nebraska, Lincoln; Nebraska Center for the Prevention of Obesity Diseases, University of Nebraska, Lincoln; Nebraska Center for Materials and Nanoscience, University of Nebraska, Lincoln; Mary and Dick Holland Regenerative Medicine Program, University of Nebraska Medical Center;

出版信息

J Vis Exp. 2022 Jun 22(184). doi: 10.3791/63420.

DOI:10.3791/63420

PMID:35815979

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10363413/

Abstract

Glioblastoma Multiforme (GBM) is the most abundant and fatal malignant brain cancer. There are more than 13,000 cases projected in the United States in 2020 and 2021. GBM tumors most often arise from astrocytes and are characterized by their invasive nature, often recruiting healthy tissues into tumor tissue. Understanding communication between astrocytes and glioblastoma cells is vital for the molecular understanding of tumor progression. This protocol demonstrates a novel patterned co-culture method to investigate contact-mediated effects of astrocytes on GBM employing layer-by-layer assembly and micro-capillary-force driven patterning. Advantages include a protein-free cell culture environment and precise control of cellular interaction dictated by the pattern dimensions. This technique provides a versatile, economical, reproducible protocol for mimicking cellular interaction between glioma and astrocytes in glioma tumors. This model can further be used to tease apart changes in GBM molecular biology due to physical contact with astrocytes or with non-contact mediated soluble cofactor communication.

摘要

多形性胶质母细胞瘤（GBM）是最常见和最致命的恶性脑癌。在美国，2020 年和 2021 年预计有超过 13000 例。GBM 肿瘤最常起源于星形胶质细胞，其特征是侵袭性，常将健康组织招募到肿瘤组织中。了解星形胶质细胞与胶质母细胞瘤细胞之间的通讯对于肿瘤进展的分子理解至关重要。本方案展示了一种新的图案共培养方法，用于通过层层组装和微毛细管力驱动的图案化来研究星形胶质细胞对 GBM 的接触介导的影响。其优点包括无蛋白的细胞培养环境以及由图案尺寸决定的精确控制细胞相互作用。该技术为模拟胶质瘤和星形胶质细胞之间在胶质瘤肿瘤中的细胞相互作用提供了一种通用、经济、可重复的方案。该模型还可以进一步用于分离由于与星形胶质细胞的物理接触或非接触介导的可溶性共因子通讯而导致的 GBM 分子生物学变化。