University of Twente Student Theses


Development of 3D neurospheroids to study immune cell infiltration in glioblastoma

Huynh, N.T. (2020) Development of 3D neurospheroids to study immune cell infiltration in glioblastoma.

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Abstract:Glioblastoma multiforme (GBM) is the most aggressive type of brain cancer with a median overall survival of just 15 months. GBM is characterized by extensive invasion and strong vascularization into adjacent brain tissue, which is responsible for the recurrence that happens within a year after primary treatment. One of the main reasons of the inefficiency of current treatments is a lack of understanding of the tumor microenvironment (TME). The TME of GBM consists of microglia, astrocytes, infiltrated macrophages and extracellular matrix (ECM), which all contribute to the GBM progression, invasion and resistance. However, knowledge about the role and recruitment of infiltrated immune cells is in particular limited. In this research, neurospheroids were developed, characterized and validated before the immune cell infiltration in GBM was studied. Spheroids were cultured from U87, HMC3 and primary astrocytes in 6 different ratios (100:0:0, 0:100:0, 0:0:100, 50:50:0, 40:0:60 and 30:30:40 respectively) using the liquid overlay method. Characterization showed that all spheroids grew in size, except for astrocytes spheroids and HMC3 spheroids, and were uniform in shape and viable without change in the spheroid composition overtime. Furthermore, the presence of astrocytes and/or HMC3 in U87 spheroids resulted in denser spheroids. Additionally, GBM-related genes (vimentin, GFAP, MMP2, TIMP1, CCL2, SSP1, CHI3L1 and GPNMB) were highly expressed in heterogeneous spheroids. However, a hypoxic core in the spheroids was not detected. When both astrocytes and microglia were included in U87 spheroids, a higher immune cell infiltration was observed. Altogether, the developed 3D neurospheroids show great potential to study the recruitment and effects of peripheral immune cells in GBM.
Item Type:Essay (Master)
Faculty:TNW: Science and Technology
Programme:Biomedical Engineering MSc (66226)
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