Main description:

The potential of human embryonic stem cells to advance not only regenerative medicine applications but also our fundamental understanding of stem cell biology continues to drive interest in research with these cells. This detailed volume collects some of the most interesting and useful protocols that have emerged in the area over the last several years. 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 expert tips on troubleshooting and avoiding known pitfalls.

Thorough and practical, Human Embryonic Stem Cell Protocols, Third Edition serves as a valuable resource to all those interested in exploring stem cell biology questions in a research setting.

Contents:

1. Derivation of Human Embryonic Stem Cell Lines from Vitrified Human Blastocysts

Cara K. Bradley, Julia Schaft , Tammie K. Roy, Biljana Dumevska, and Teija T. Peura

2. Characterizing Pluripotent Stem Cells Using the TaqMan (R) hPSC ScorecardTM Panel

Jeffrey Fergus, Rene Quintanilla, and Uma Lakshmipathy

3. Growth of Human Pluripotent Stem Cells Using Functional Human Extracellular Matrix

Andres Sanz-Garcia, Miodrag Stojkovic, and Carmen Escobedo-Lucea

4. Efficient Expansion of Dissociated Human Pluripotent Stem Cells Using a Synthetic Substrate

Eihachiro Kawase

5. Microarray Approach to Identify the Signaling Network Responsible for Self-Renewal of Human Embryonic Stem Cells

Noboru Sato and Ali Brivanlou

6. Monitoring Stemness in Long-Term hESC Cultures by Real-Time PCR

Amparo Galan and Carlos Simon

7. Study of Gap Junctions in Human Embryonic Stem Cells

Alice Pebay and Raymond C.B. Wong

8. Immunofluorescence Microscopy and mRNA Analysis of Human Embryonic Stem Cells (hESCs) Grown Under Feeder-Free Conditions

Maj Linea Vestergaard, Aashir Awan, Caroline Becker Warzecha, Soren Tvorup Christensen, and Claus Yding Andersen

9. Analysis of Intracellular Calcium Signaling in Human Embryonic Stem Cells

Adrienn Pentek, Katalin Paszty, and Agota Apati

10. Genetic Manipulation of Human Embryonic Stem Cells

Rachel Eiges

11. Genetic Modification in Human Pluripotent Stem Cells by Homologous Recombination and CRISPR/Cas9 System

Haipeng Xue, Jianbo Wu, Shenglan Li, Mahendra S. Rao, and Ying Liu

12. Use of Multicolor Flow Cytometry for Isolation of Specific Cell Populations Deriving from Differentiated Human Embryonic Stem Cells

Isabella Mengarelli, Andrew Fryga, and Tiziano Barberi

13. Definitive Endoderm Differentiation of Human Embryonic Stem Cells Combined with Selective Elimination of Undifferentiated Cells by Methionine Deprivation

Tomonori Tsuyama, Nobuaki Shiraki, and Shoen Kume

14. Derivation of Endothelial Cells and Pericytes from Human Pluripotent Stem Cells

Sravanti Kusuma and Sharon Gerecht

15. Differentiation of Human Embryonic Stem Cells on Periodontal Ligament Fibroblasts

Y. Murat Elcin, Bulend Inanc, and A. Eser Elcin

16. Derivation of Epithelial Cells from Human Embryonic Stem Cells as an In Vitro Model of Vocal Mucosa

Vlasta Lungova, Ciara Leydon, and Susan Thibeault

17. Human Embryonic and Hepatic Stem Cell Differentiation Visualized in Two and Three Dimensions Based on Serial Sections

Peter S. Vestentoft, Christian B. Brochner, Niels Lynnerup, Claus Yding Andersen, and Kjeld Mollgard

18. Derivation of Chondrogenic Cells from Human Embryonic Stem Cells for Cartilage Tissue Engineering

Wei Seong and Tong Cao

19. Microgrooved Surface Modulates Neuron Differentiation in Human Embryonic Stem Cells

David Lu, Chi-Shuo Chen, Chao-Sung Lai, Sushant Soni, Taranze Lam, Clerance Le, Eric Y-T Chen, Thien Nguyen, and Wei-Chun Chin

20. Directed Differentiation of Human Embryonic Stem Cells into Neural Progenitors

Erin Banda and Laura Grabel

21. Direct Conversion of Pluripotent Human Embryonic Stem Cells Under Defined Culture Conditions into Human Neuronal or Cardiomyocyte Cell Therapy Derivatives

Xuejun H. Parsons

22. Generation of Epithelial Cell Populations from Human Pluripotent Stem Cells Using a Small Molecule Inhibitor of Src Family Kinases

Joshua A. Selekman, Xiaojun Lian, and Sean P. Palecek

23. Dual-SMAD Inhibition/WNT Activation-Based Methods to Induce Neural Crest and Derivatives from Human Pluripotent Stem Cells

Stuart M. Chambers, Yvonne Mica, Gabsang Lee, Lorenz Studer, and Mark J. Tomishima

24. Accelerated Three-Dimensional Neuroepithelium Formation from Human Embryonic Stem Cells and Its Use for Quantitative Differentiation to Human Retinal Pigment Epithelium

Yu Zhu, Sven Schreiter, and Elly M. Tanaka

25. Efficient Production of Photoreceptor Precursor Cells from Human Embryonic Stem Cells

Anat Yanai, Christopher Laver, Aaron W. Joe, and Kevin Gregory-Evans

26. Generation of Megakaryocytes and Platelets from Human Pluripotent Stem Cells

Marjorie Pick

27. A Simple Protocol for the Generation of Cardiomyocytes from Human Pluripotent Stem Cells

Glen Lester Sequiera, Ashish Mehta, and Winston Shim