Human Embryonic Stem Cell Protocols (3^rd Ed., Softcover reprint of the original 3rd ed. 2015) Methods in Molecular Biology Series, Vol. 1307

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.

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^® hPSC Scorecard^TM 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 Galán and Carlos Simón

7. Study of Gap Junctions in Human Embryonic Stem Cells

            Alice Pébay 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, Søren Tvorup Christensen, and Claus Yding Andersen

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

            Adrienn Péntek, Katalin Pászty, and Ágota Apáti

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 Elçin, Bülend İnanç, and A. Eser Elçin

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. Brøchner, Niels Lynnerup, Claus Yding Andersen, and Kjeld Møllgård

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

Includes cutting-edge methods and protocols involving human embryonic stem cells

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts