This volume explores the fields of stem cell biology, regenerative medicine, and cancer biology. The chapters in this book cover topics such as embryonic stem cells (ES) and induced pluripotent cells (iPS); ES/iPSC derived tissue stem cells; tissue resident stem cells; assays for primitive malignant cells; and CRISPR/Cas9 engineering and transgenic model development. 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 tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and thorough, Stem Cell Assays: Methods and Protocols is a valuable laboratory manual that allows researchers and scientists to learn about new techniques and approaches in this developing field.
1. Rapid and Highly Efficient Isolation and Purification of Human Induced Pluripotent Stem Cells Xiugong Gao, Robert L. Sprando, and Jeffrey J. Yourick
2. Artificial Activation of Murine Oocytes using Strontium to Derive Haploid and Diploid Parthenotes Daphne Norma Crasta, Satish Kumar Adiga, Nagarajan Kannan, and Guruprasad Kalthur
3. Generation of Human iPSC from Small Volume Peripheral Blood Samples Doreen Kloos and Nico Lachmann
4. Distinguishing between Endodermal and Pluripotent Stem Cell Lines during Somatic Cell Reprogramming A. Moauro and A. Ralston
5. Measuring Early Germ-Layer Specification Bias in Human Pluripotent Stem Cells Alexander Keller, Nusa Krivec, Christina Markouli, and Claudia Spits
6. Detection of Soluble and Insoluble Protein Species in Patient-Derived iPSCs Stephanie Santarriaga, Ian Luecke, and Allison D. Ebert
7. Extracellular Flux Analysis of Mitochondrial Function in Pluripotent Stem Cells Enkhtuul Tsogtbaatar, Katherine Minter-Dykhouse, Alicia Saarinen, and Clifford D. L. Folmes
8. Assessment of Endothelial-to-Hematopoietic Transition of Individual Hemogenic Endothelium and Bulk Populations in Defined Conditions Gene I. Uenishi, Ho Sun Jung, and Igor I. Slukvin
Part II ES/iPS Derived Tissue Stem Cells
9. Homogenous Differentiation of Functional Hepatocytes from Human Induced Pluripotent Stem Cells Rong Li, Yang Zhao, Jeffrey J. Yourick, Robert L. Sprando, and Xiugong Gao
10. Differentiation of Human Induced Pluripotent Stem Cells into Cortical Neurons to Advance Precision Medicine M. Catarina Silva, Ghata Nandi, and Stephen J. Haggarty
11. Differentiation of iPS-Cells into Peripheral Sensory Neurons Anika Neureiter, Esther Eberhardt, and Angelika Lampert
12. Culture of Human iPSC-Derived Motoneurons in Compartmentalized Microfluidic Devices and Quantitative Assays for Studying Axonal Phenotypes Maria Giovanna Garone, Chiara D’Antoni, and Alessandro Rosa
13. iPS Cell-Differentiation into Brain Microvascular Endothelial Cells Angelica Medina and Hengli Tang
14. Chromatin Immunoprecipitation in Human Pluripotent Stem Cell-Derived 3D Organoids to Analyze DNA-Protein Interactions Wei Xuan Tan, Chek Mei Bok, Natasha Hui Jin Ng, and Adrian Kee Keong Teo
15. Generation of Embryonic Origin-Specific Vascular Smooth Muscle Cells from Human Induced Pluripotent Stem Cells Mengcheng Shen, Chun Liu, and Joseph C. Wu
16. Generation of Salivary Gland Organoids from Mouse Embryonic Stem Cells Junichi Tanaka and Kenji Mishima
17. In Vitro Generation of Heart Field Specific Cardiomyocytes Arash Pezhouman, Ngoc B. Nguyen, Allison Shevtsov, Rong Qiao, and Reza Ardehali
Part III Tissue Stem Cells
18. Isolation and Characterization of Extracellular Vesicles Derived from Human Umbilical Cord Mesenchymal Stem Cells Noridzzaida Ridzuan, Darius Widera, and Badrul Hisham Yahaya
19. Identification and Validation of CRISPR/Cas9 Off-Target Activity in Hematopoietic Stem and Progenitor Cells So Hyun Park, Ciaran M. Lee, and Gang Bao
20. Genome Engineering of Hematopoietic Stem Cells using CRISPR/Cas9 System Nivedhitha Devaraju, Vignesh Rajendiran, Nithin Sam Ravi, and Kumarasamypet M. Moahnkumar
21. Generation of Rat Neural Stem Cells to Produce Different Astrocyte Phenotypes Rebecca Sherrard Smith, Susan C. Barnett, and Susan L. Lindsay
22. In Situ Quantification and Isolation of Müller Glial Cells by Fluorescence Activated Cell Sorting from the Regenerating Larval Zebrafish Retina Jeffrey Stulberg and Vincent Tropepe
23. Quantification and Cloncal Culture of Neural Stem Cells from the Hippocampus of Adult Mouse Yoko Amagase, Hiroko Izumi-Nakaseko, Atsushi Sugiyama, and Yoshinori Takei
24. Reprogramming Mouse Oviduct Epithelial Cells using In Vivo Electroporation and CRISPR/Cas9 Mediated Genetic Manipulation Matthew J. Ford and Yojiro Yamanaka
25. Generation of Human Liver Chimeric Mice and Harvesting of Human Hepatocytes from Mouse Livers Rui Wei, Chi-Wa Cheng, Wai-In Ho, Kwong-Man Ng, Miguel A. Esteban, and Hung-Fat Tse
26. Application of 3D Culture Assays to Study Breast Morphogenesis, Epithelial Plasticity, and Cellular Interactions in an Epithelial Progenitor Cell Line Anna Karen Sigurdardottir, Bylgja Hilmarsdottir, Thorarinn Gudjonsoon, and Gunnhildur Asta Traustadottir
27. A Unified Protocol to Streamline Molecular and Cellular Analysis for Three-Dimensional Cell Cultures Lisa M. Kim, Paul Y. Kim, and Cheuk Y. Leung
28. Mesenchymal Stem Cell Seeding on 3D Scaffolds Agata Kurzyk
29. Assaying Candidate Human Skin Keratinocyte Stem Cells by Determining their Long-Term Serial Proliferative Output in Culture Zalitha Pieterse and Pritinder Kaur
30. Protocol for the Detection of Organoid-Initiating Cell Activity in Patience-Derived Single Fallopian Tube Epithelial Cells Liang Feng, Wenmei Yang, Hui Zhao, Jamie Bakkum-Gamez, Mark E. Sherman, and Nagarajan Kannan
31. Quantification of Mucle Stem Cell Differentiation using Live-Cell Imaging and Eccentricity Measures Paige C. Arneson-Wissink and Jason D. Doles
Part IV Malignancy
32. The Enrichment of Breast Cancer Stem Cells from MCF7 Breast Cancer Cell Line using Spheroid Culture Technique Anan A. Ishtiah and Badrul Hisham Yahaya
33. Assessment of Breast Cancer Stem Cell Activity using a Spheroid Formation Assay Ajeya Nandi and Rumela Chakrabarti
34. Enrichment of Cancer Stem Cells in a Tumor Sphere Assay Abhijeet P. Deshmukh, Petra den Hollander, Nick Kuburich, Suhas Vasaikar, Robiya Joseph, and Sendurai A. Mani
35. In Vitro Quantification of Cancer Stem Cells using a Mammosphere Formation Assay Nick Kuburich, Petra den Hollander, Abhijeet P. Deshmukh, Suhas Vasaikar, Robiya Joseph, Max S. Wicha, and Sendurai A. Mani
36. Designing Genetically Engineered Mouse Models (GEMMSs) using CRISPS Mediated Genome Editing Jade Desjardins, Mitra Cowan, and Yojiro Yamanaka
37. Assays for the Spectrum of Circulating Tumor Cells Xuanmao Jiao, Chandan Upadhyaya, Zhao Zhang, Jun Zhao, Zhiping Li, Vivek I. Patel, and Richard G. Pestell
38. Limiting Dilution Tumor Initiation Assay: An In Vitro Approach for the Study of Cancer Stem Cells Petra den Hollander, Robiya Joseph, Suhas Vasaikar, Nick Kuburich, Abhijeet P. Deshmukh, and Sendurai A. Mani
39. Orthotopic Xenografts of Colorectal Cancer Stem Cells Maria Laura De Angelis, Federica Francescangeli, Ann Zeuner, and Marta Baiocchi
Subject Index List…
This volume explores the fields of stem cell biology, regenerative medicine, and cancer biology. The chapters in this book cover topics such as embryonic stem cells (ES) and induced pluripotent cells (iPS); ES/iPSC derived tissue stem cells; tissue resident stem cells; assays for primitive malignant cells; and CRISPR/Cas9 engineering and transgenic model development. 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 tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and thorough, Stem Cell Assays: Methods and Protocols is a valuable laboratory manual that allows researchers and scientists to learn about new techniques and approaches in this developing field.