Abstract: Glioblastoma stem cells (GSC) are cells in glioblastoma (GBM) with self-renewal capabilities and ability to initiate tumours and differentiate into multiple lineages, contributing to tumour heterogeneity and poor prognosis of GBM. Mitophagy is a type of autophagy that targets mitochondria, removing unhealthy or unnecessary mitochondria. Tumour microenvironment (TME) is the microenvironment that surrounds the tumour, which plays an important role in various aspects of the tumour, such as tumour behaviour, proliferation and death. The effects of mitophagy in the TME on GSC have not been studied extensively in current literature. Mitophagy activation in the TME has been discovered in our study to cause significant changes in gene expression and reduce tumorigenicity of GSCs. This study may provide insight into alternative treatment possibilities and pathophysiology of GBM.