Stem cells are self-replicating and undifferentiated, meaning their function is not yet cell, tissue, or organ-specific. Due to the unique nature of these cells, research into their biology and function holds great promise for therapeutic applications through replacement or repair of diseased and damaged cells. This reader-friendly manual provides a practical "hands on" guide to the culture of human embryonic and somatic stem cells. By presenting methods for embryonic and adult lines side-by-side, the authors lay out an elegant and unique path to understanding the science of stem cell practice. The authors begin with a broad-based introduction to the field, and also review legal and regulatory issues and patents. Each experimental strategy is presented with an historical introduction, detailed method, discussion of alternative methods, and common pitfalls. This lab guide for researchers also serves as a textbook for undergraduate and graduate students in laboratory courses.
Scientists engaged in any aspect of stem cell research, as well as undergraduate and graduate students enrolled in laboratory courses.
Foreword by James Battey
Foreword by Jeannie Fontana
Part I: Basic methods: Maintenance of stem cell cultures
1.Human Embryonic Stem Cell Culture
2.Feeder layers and feeder-free culture 3.Mouse Embryonic Fibroblast Feeder Cells 4.Cryopreservation of hESC
Part II: Basic methods: Characterization of stem cells 5.Classical Cytogenetics: Karyotyping 6.Spectral Karyotyping and Fluorescent in situ Hybridization 7.Using SNP Genotyping to Identify Cell Lines, Determine Genomic Abnormalities and Monitor DNA Methylation Patterns 8.FACS/Cell sorting 9.Fluorescence Immunocytochemical Analysis of Stem Cells 10.Characterization of Stem Cells Using RT-PCR 11.Gene Expression Profiling of Stem Cells by Microarray 12.Teratomas produced from Human Embryonic Stem Cells Xenografted into Immunodeficient Mice 13.Generation of HESC-Derived Teratomas Part III: Differentiation of stem cells 14.“Embryoid Body” Formation and Neuroepithelia Differentiation 15.Motor Neuron and Dopamine Neuron Differentiation 16.Oligodendrocyte Differentiation from hESC 17.Cardiac 18.Hematopoietic Part IV: Genetic manipulation of stem cells 19.Genetic Manipulation of hESC: Lentivirus Vectors 20.Methods developed for mouse ES cells Part V: Advanced methods 21.Derivation of human ES cells 22.Neural Stem Cell Culture 23.Transplantation of stem cells 24.In vitro fertilization 25.Functional characterization-neural cells Part VI: Practical issues in establishing a human ES cell laboratory 26.Setting Up a Facility for hESC Research
- No. of pages:
- © Academic Press 2007
- 1st June 2007
- Academic Press
- eBook ISBN:
- Hardcover ISBN:
The Scripps Research Institute, La Jolla, CA
The Burnham Institute, La Jolla, California, U.S.A.