This comprehensive laboratory manual provides detailed, step-by-step protocols for maintaining human induced pluripotent stem cells (hiPSCs), directing their differentiation into unguided cerebral organoids, and performing essential downstream analyses. Written for researchers, graduate students, and laboratory technicians, this guide bridges the gap between theoretical knowledge and practical application in 3D brain modelling. Each protocol includes troubleshooting tips and best practices derived from established methodologies. Whether you are establishing organoid culture in your laboratory for the first time or refining existing techniques, this manual serves as an indispensable bench-side companion for neurodevelopmental research, disease modelling, and drug screening applications.