This is the third of three planned volumes in the "Methods in Enzymology" series on the topic of stem cells. This volume is a unique anthology of stem cell techniques written by experts from the top laboratories in the world. The contributors not only have hands-on experience in the field but often have developed the original approaches that they share with great attention to detail. The chapters provide a brief review of each field followed by a 'cookbook' and handy illustrations. The collection of protocols includes the isolation and maintenance of stem cells from various species using 'conventional' and novel methods, such as derivation of ES cells from single blastomeres, differentiation of stem cells into specific tissue types, isolation and maintenance of somatic stem cells, stem cell-specific techniques and approaches to tissue engineering using stem cell derivatives. The reader will find that some of the topics are covered by more than one group of authors and complement each other.
Comprehensive step-by-step protocols and informative illustrations can be easily followed by even the least experienced researchers in the field, and allow the setup and troubleshooting of these state-of-the-art technologies in other laboratories. The book provides complete coverage spanning from derivation/isolation of stem cells, and including differentiation protocols, characterization and maintenance of derivatives and tissue engineering. It presents the latest and most innovative technologies. It addresses therapeutic relevance including FDA compliance and tissue engineering.
Section I. In vitro experimentation Human embryo culture. Characterization and evaluation of human embryonic stem cells. Feeder-free culture of human embryonic stem cells. Transgene Expression and RNA Interference in Embryonic Stem Cells. Lentiviral vectors-mediated Gene Delivery into Human Embryonic Stem Cells. Engineering embryonic stem cells with recombinase systems. Gene trapping in embryonic stem cells. Genechips in stem cell research. Microarray analysis of stem cells and differentiation. Purification and hematopoietic stem cells using the side population. Cellular reprogramming. Section II. Tissue engineering and regenerative medicine Tissue engineering using adult stem cells. Tissue Engineering using embryonic stem cells. Engineering cardiac tissue from embryonic stem cells. Mesenchymal Stem Cell and Tissue Engineering. Bone reconstruction with bone marrow stromal cells. Engineering three-dimensional tissue structures using Stem Cells. Immunogenicity of embryonic stem cells and their derivatives. Clinical uses of therapies derived from stem cells.