In this fully revised edition of an established classic, expert researchers and clinicians describe in step-by-step detail updated techniques for the isolation and growth of the major primary cell types, such as kidney proximal tubule cells, hepatocytes, keratinocytes, and cardiomyocytes. The authors offer readily reproducible new methods for the differentiation of embryonic stem (ES) cells into various hematopoietic cell types, for fetal thymic organ culture, and for the isolation and culture of specialized cell types, such as mammary progenitor cells, skeletal muscle myofibers, mesenchymal cells, neural stem cells, hematopoietic cells, stromal cell lines, and endothelial cells. Additional chapters describe new techniques (leukocyte rolling, isolation of side population cells, and scalable production of ES-derived cells) and detail quality control methods for cell lines (detection and elimination of mycoplasma, DNA fingerprinting, and cytogenetic analysis). The protocols follow the successful Methods in Molecular Biology(TM) series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Up-to-date and highly practical, Basic Cell Culture Protocols, Third Edition, offers basic scientists and clinician-researchers powerful tools to isolate, culture, and characterize the promising specialized cell types in demand today.