As part of the boom in biotechnology, particularly in tissue engineering, primary human cell culture has become a major pillar in academic research and the biopharmaceutical industry. Obtaining a viable culture from a tissue sample and maintaining it for experimental, diagnostic or therapeutic purposes can be quite a challenge. Based on laboratory protocols and practical experience from many years of primary cell culture, this manual presents the basic steps necessary for culturing primary human cells. Written by students for students, the manual serves well as a practical guide to primary human cell culture. The authors have left much space for notes and the design of the manual is such that it can be continuously upgraded and extended. The content of this manual is by no means exhaustive. Protocols for specific cell types, out of over 200 different cell types in the human body, were selected from major tissue groupings in the body. They should serve as a foundation for individual researchers to experiment, explore, and establish niche protocols for their specific needs. Inspired by the practical clinical checklists available to residents and trainees in medicine, the authors have chosen a compact physical format that can fit into the pocket of a lab coat.