Human pluripotent stem cells (hPSCs ) have the unique potential to form every cell type in the body. This potential provides opportunities for generating human progenitors and other differentiated cell types for understanding human development and for use in cell type-specific therapies. Equally important is the ability to engineer stem cells and their derived progenitors to mimic specific disease models. This chapter will focus on the propagation and characterization of human neural progenitors (hNPs ) derived from hPSCs with a particular focus on engineering hNPs to generate in vitro disease models for human neuro-mitochondrial disorders. We will discuss the methodologies for culturing and characterizing hPSCs and hNPs; and protocols for engineering hNPs by using a novel mitochondrial transfection technology.