Human Cell Culture Protocols

This third edition of Human Cell Culture Protocols expands upon the previous editions with current, detailed protocols for the isolation and culture of a range of primary cells from human tissues. It features cutting-edge methods ready for the laboratory.

Human cell culture is not a new topic, but the development of new molecular techniques and reagents which can be used to investigate cell function and the responsible intracellular mechanisms make it a continuing requirement. This third edition of Human Cell Culture Protocols expands upon the previous editions with current, detailed protocols for the isolation and culture of a range of primary cells from human tissues. With new chapters on pancreatic cells needed for basic studies on the pathogenesis of diabetes and for their application for islet transplantation, the book also delves into protocols for hepatocytes, skin cells, lung cells, parathyroid cells, gastric cells, renal cells, adipocytes, ovarian cells, bone cells, vascular smooth muscle cells, vascular endothelial cells, regulatory T cells, blood mononuclear cells, as well as new techniques being applied to human cell culture, particularly the use of biocompatible scaffolds to grow cells, the in vitro use of laser microdissection to isolate cells from culture, and automated cell culture. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Human Cell Culture Protocols, Third Edition makes it possible for a worker with basic cell culture training, whether in the fields of cell biology, gene therapy, and cell transplantation, to prepare cell cultures of the specific cell type necessary to forward their vital research.

Supplies techniques that can be adapted to obtain cell cultures from diseased cells to gain understanding of the pathogenic mechanisms involved Features cutting-edge methods ready for the laboratory Includes expert tips and implementation advice to ensure successful results

Inhalt

Introduction to Cell Culture.- Isolation and Cultivation of Dermal Stem Cells that Differentiate into Functional Epidermal Melanocytes.- Isolation, Cultivation, and Application of Human Alveolar Epithelial Cells.- Culture of Parathyroid Cells.- Functional Analysis of Human Islets of Langerhans Maintained in Culture.- Conversion of Non-Endocrine Human Pancreatic Cells to Insulin Producing Cells for Treatment of Diabetes.- Evaluation of Cytochrome P450 Activities in Human Hepatocytes In Vitro.- Human Hepatocytes: Isolation, Culture, and Quality Procedures.- Preclinical Testing of Virotherapeutics for Primary and Secondary Tumors of the Liver.- Isolation and Functional Studies of Human Fetal Gastric Epithelium in Primary Culture.- Isolation, Characterization, and Culture of Normal Human Intestinal Crypt and Villus Cells.- Primary Culture of Human Renal Proximal Tubule Epithelial Cells and Interstitial Fibroblasts.- Glomerular Epithelial and Mesangial Cell Culture and Characterization.- Isolation and Culture of Human Adipocytes and Adipose Tissue.- Primary Culture of Human Adipocyte Precursor Cells: Expansion and Differentiation.- Primary Culture of Ovarian Cells for Research on Cell Interactions in the Hormonal Control of Steroidogenesis.- Human Vascular Smooth Muscle Cell Culture.- Culture of Human Endothelial Cells from Umbilical Veins.- Human Peripheral Blood Mononuclear Cell Culture for Flowcytometric Analysis of Phosphorylated Mitogen-Activated Protein Kinases.- Human CD4+CD25 high CD127 low/neg Regulatory T Cells.- Human Chondrocyte Cultures as Models of Cartilage-Specific Gene Regulation.- Isolation and Culture of Human Osteoblasts.- Human Osteoclast Culture and PhenotypicCharacterization.- Effects of Temperature Generated from the Holmium:YAG Laser on Human Osteoblasts in Monolayer Tissue Culture.- Laser Microdissection Microscopy: Application to Cell Culture.- Automated Adherent Human Cell Culture (Mesenchymal Stem Cells).- Culturing and Differentiating Human Mesenchymal Stem Cells for Biocompatible Scaffolds in Regenerative Medicine.