Glioblastoma (GBM) is the most prevalent and malignant primary brain tumor, characterized by the presence of self-renewing, tumorigenic cancer stem cells (CSCs). These CSCs contribute to tumor initiation and therapeutic resistance. The development of GBM recapitulates the cellular hierarchies found in normal tissues, with CSCs at the apex of the hierarchy. The molecular mechanisms governing CSCs have informed the development of targeted therapeutics for GBM. CSCs function within an ecological system, actively remodeling the microenvironment and receiving maintenance cues from their niches. The complexity of GBM, including genetic, epigenetic, and metabolic factors, has been extensively studied, but challenges remain in distinguishing CSCs from their differentiated progeny. Markers such as CD133, CD44, and integrin α6 have been used to enrich CSCs, but their utility is limited by the lack of definitive markers. Intrinsic regulators of CSCs include genetics, epigenetics, and metabolism, while extrinsic regulators involve the microenvironment, immune system, and niche factors. CSCs exhibit plasticity in metabolic pathways and are dependent on specific signaling pathways like Notch, BMP, NF-κB, and Wnt. Therapeutic targeting of CSCs is challenging due to the lack of universally informative markers and the shared molecular pathways between CSCs and normal stem cells. Mathematical modeling approaches have been used to understand the dynamics of CSCs and GBM growth, providing insights into key pathways essential for self-renewal and therapeutic responses. Future research aims to refine the understanding of CSCs and develop more effective therapies.Glioblastoma (GBM) is the most prevalent and malignant primary brain tumor, characterized by the presence of self-renewing, tumorigenic cancer stem cells (CSCs). These CSCs contribute to tumor initiation and therapeutic resistance. The development of GBM recapitulates the cellular hierarchies found in normal tissues, with CSCs at the apex of the hierarchy. The molecular mechanisms governing CSCs have informed the development of targeted therapeutics for GBM. CSCs function within an ecological system, actively remodeling the microenvironment and receiving maintenance cues from their niches. The complexity of GBM, including genetic, epigenetic, and metabolic factors, has been extensively studied, but challenges remain in distinguishing CSCs from their differentiated progeny. Markers such as CD133, CD44, and integrin α6 have been used to enrich CSCs, but their utility is limited by the lack of definitive markers. Intrinsic regulators of CSCs include genetics, epigenetics, and metabolism, while extrinsic regulators involve the microenvironment, immune system, and niche factors. CSCs exhibit plasticity in metabolic pathways and are dependent on specific signaling pathways like Notch, BMP, NF-κB, and Wnt. Therapeutic targeting of CSCs is challenging due to the lack of universally informative markers and the shared molecular pathways between CSCs and normal stem cells. Mathematical modeling approaches have been used to understand the dynamics of CSCs and GBM growth, providing insights into key pathways essential for self-renewal and therapeutic responses. Future research aims to refine the understanding of CSCs and develop more effective therapies.