Basic Cell Culture Protocols

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.

Culture of Primary Adherent Cells and a Continuously Growing Nonadherent Cell Line. - Detection of Mycoplasma Contaminations. - Eradication of Mycoplasma Contaminations. - Authentication of Scientific Human Cell Lines. - Cytogenetic Analysis of Cell Lines. - Human and Mouse Hematopoietic Colony-Forming Cell Assays. - Isolation and Culture of Murine Macrophages. - Isolation and Culture of Human Macrophages. - Development of T-Lymphocytes in Mouse Fetal Thymus Organ Culture. - In Vitro Generation of Lymphocytes From Embryonic Stem Cells. - Hematopoietic Development of Human Embryonic Stem Cells in Culture. - Generation of Murine Stromal Cell Lines. - Culture of Human and Mouse Mesenchymal Cells. - Isolation, Purification, and Cultivation of Murine and Human Keratinocytes. - Isolation and Culture of Primary Human Hepatocytes. - Primary Kidney Proximal Tubule Cells. - Enzymatic Dissociation and Culture of Normal Human Mammary Tissue to Detect Progenitor Activity. - Generation and Differentiation of Neurospheres From Murine Embryonic Day 14 Central Nervous System Tissue. - Isolation and Culture of Skeletal Muscle Myofibers as a Means to Analyze Satellite Cells. - Adult Ventricular Cardiomyocytes. - Isolation and Culture of Primary Endothelial Cells. - Studying Leukocyte Rolling and Adhesion In Vitro Under Flow Conditions. - Isolation and Characterization of Side Population Cells. - Scalable Production of Embryonic Stem Cell-Derived Cells.