A chemically defined medium (E8) and vitronectin-coated surfaces were developed for the derivation and culture of human induced pluripotent stem (iPS) cells. The E8 medium contains eight components, including DMEM/F12, insulin, selenium, transferrin, L-ascorbic acid, FGF2, and TGFβ or NODAL. This medium supports the undifferentiated proliferation of human embryonic stem (ES) and iPS cells, maintains pluripotency markers and normal karyotypes, and allows for efficient reprogramming of fibroblasts into iPS cells. The vitronectin-coated surfaces enhance cell attachment and survival in E8 medium. The E8 medium improves reprogramming efficiency compared to other methods, and is suitable for most common cell culture practices. The study demonstrates that the E8 medium supports the derivation of integration-free iPS cells from patient-derived fibroblasts, and that the use of defined conditions improves the consistency and reliability of stem cell culture. The study also highlights the importance of removing serum albumin and other animal-derived components to achieve a fully defined culture system. The E8 medium is cost-effective, simplifies quality control, and supports long-term culture of ES and iPS cells. The study provides a standardized, chemically defined system for the derivation and culture of human iPS cells, which should facilitate both research and clinical applications.A chemically defined medium (E8) and vitronectin-coated surfaces were developed for the derivation and culture of human induced pluripotent stem (iPS) cells. The E8 medium contains eight components, including DMEM/F12, insulin, selenium, transferrin, L-ascorbic acid, FGF2, and TGFβ or NODAL. This medium supports the undifferentiated proliferation of human embryonic stem (ES) and iPS cells, maintains pluripotency markers and normal karyotypes, and allows for efficient reprogramming of fibroblasts into iPS cells. The vitronectin-coated surfaces enhance cell attachment and survival in E8 medium. The E8 medium improves reprogramming efficiency compared to other methods, and is suitable for most common cell culture practices. The study demonstrates that the E8 medium supports the derivation of integration-free iPS cells from patient-derived fibroblasts, and that the use of defined conditions improves the consistency and reliability of stem cell culture. The study also highlights the importance of removing serum albumin and other animal-derived components to achieve a fully defined culture system. The E8 medium is cost-effective, simplifies quality control, and supports long-term culture of ES and iPS cells. The study provides a standardized, chemically defined system for the derivation and culture of human iPS cells, which should facilitate both research and clinical applications.