Full Description
This manual is a comprehensive compilation of "methods that work" for deriving, characterizing, and differentiating hPSCs, written by the researchers who developed and tested the methods and use them every day in their laboratories. The manual is much more than a collection of recipes; it is intended to spark the interest of scientists in areas of stem cell biology that they may not have considered to be important to their work. The second edition of the Human Stem Cell Manual is an extraordinary laboratory guide for both experienced stem cell researchers and those just beginning to use stem cells in their work.
Contents
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
Part VII: Essays on stem cell biology
27.Intellectual Property: Owning the Stem Cell
28.Ethical Concerns for Stem Cell Research
29.Guidelines for hESC Research Oversight (ESCRO) Committees
