This manual is designed to serve as a practical guide to primary human cell culture, which is integral in both academic and industrial biotechnology research. As in the first edition, the content of the manual is not exhaustive, but rather contains selected protocols for specific cell types from major tissue groupings in the body. This improved second edition also includes a new section on stem cells and additional material on transfection. It should serve as a foundation for individual researchers to experiment, explore, and establish niche protocols for their specific needs. With its compact physical format that makes it portable and flexible for usage in a laboratory setting, the manual will be a useful guide for all beginners in primary human cell culture work.