In recent years, human pluripotent stem cells such as human embryonic stem (ES) cells and human inducible pluripotent stem (iPS) cells have been widely used not only in basic research but also in drug discovery research and application to regenerative medicine. Simultaneously, because the research environment including supply of cultivation materials has been improved, culture of human pluripotent stem cells can be easily carried out, and the number of researchers using human pluripotent stem cells is increasing. On the other hand, it is important points for researchers/engineers to share the basic concepts of technology and its background and to ensure the reproducibility of the research results. Characters of human pluripotent stem cells are significantly different from that of somatic cells that have been used in the past, and there are many points to keep in mind even if researchers/engineers have a vast experience in culturing mammalian cells. Therefore, we hope that human pluripotent stem cells will be usefully utilized and drew a draft guidance of “key points of consideration for pluripotent stem cell culture”. This draft describes the contents to be confirmed at the start of use of human pluripotent stem cells in seven sections (agreed with laws and regulations/guidelines/MTA, diversity of pluripotent stem cell, culture material, thawing operation, medium exchange and passage, freezing operation, and culture management). We propose here the guidance as the concept shared by researchers and engineers who carry out cell culture of pluripotent stem cells, and hope that utilization and reliability of research using pluripotent stem cells will improve.