This volume presents up-to-date methods that allow primary stem cells from a variety of sources to be isolated, cultured in vitro, detected and measured for specific applications. These applications range from those in basic, stem cell and veterinary research to toxicology, cellular therapy and regenerative medicine. There is a slight bias towards the blood-forming system as more is known about the blood-forming or hematopoietic system than any other primary stem cell system. These unique properties and characteristics are discussed and examined, mostly at the cellular level and in detail in this book. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.
Authoritative and accessible, Stem Cell Protocols provides novices with the fundamentals necessary to develop new technologies necessary for basic and clinical research in the future, and will aid professionals in finding new methodologies to provide a wider viewpoint and an even greater scope for their own research.
TOC:
1. A Short Primer in Stem Cell Biology
Ivan N. Rich
2. Measurement of Hematopoietic Stem Cell Proliferation, Self-Renewal and Expansion Potential
Ivan N. Rich
3. Measuring the Aging Process in Stem Cells
Yi Liu, Gary Van Zant, and Ying Liang
4. Measuring the Potency of a Stem Cell Therapeutic
Holli Harper and Ivan N. Rich
5. Culturing Protocols for Human Multipotent Adult Stem Cells
Bart Vaes, Sara Walbers, Kristel Gijbels, David Craeye, Robert Deans, Jef Pinxteren, and Wouter van’t Hof
6. The Isolation of Murine Bone Marrow Scavenging Sinusoidal Endothelial Cells
Peter A. M. McCourt, Ana Oteiza, and Susan K. Nilsson
7. Analysis of Circadian Rhythms in Embryonic Stem Cells
Jiffin K. Paulose, Edmund B. Rucker III, Vincent M. Cassone
8. Measuring Stem Cell Circadian Rhythms
William Hrushesky and Ivan N. Rich
9. Cryopreservation of Human Pluripotent Stem Cells: A General Protocol
Tamamichi Miyazaki and Hirofumi Suemori
10. Biological Differences Between Native and Cultured Mesenchymal Stem Cells – Implications for Therapies
Elena Jones and Richard Schaefer
11. The Use of Multiparameter Flow Cytometry and Cell Sorting to Characterize Native Human Bone Marrow Mesenchymal Stem Cells (MSC)
Sally Boxall and Elena Jones
12. High Yield Recovery of Equine MEsenchymal Stem Cells from Umbilical Cord Matrix/Wharton’s Jelly Using a Semi-Automated Process
Timo Z. Nazari-Shafti, Ivone G. Bruno, Rudy Martinez, Michael E. Coleman, Eckhard U. Alt, and Scott R. McClure
13. Isolation and Functional Assessment of Cutaneous Stem Cells
Yanne S. Doucet and David M. Owens
14. Isolation of Adult Stem Cell Populations from the Human Cornea
Matthew J. Branch, Wing-Yan Yu, Carl Sheridan, and Andrew Hopkinson
15. Advanced Imaging and Tissue Engineering of the Human Limbal Epithelial Stem Cell Niche
Isobel Massie, Marc Dziasko, Alvena Kureshi, Hannah J. Levis, Louise Morgan, Michael Neale, Radhika Sheth, Victoria E. Tovell, Amanda J. Vernon, James L. Funderburgh, and Jule T. Daniels
16. Isolation and Characterization of Stem Cells in the Adult Mammalian Ovary
Seema Parte, Hiren Patel, Kalpana Sriraman, and Deepa Bhartiya
17. Clonal Culture of Adult Mouse Lung Epithelial Stem/Progenitor Cells
Jonathan L. McQualter and Ivan Bertoncello
18. Culture and Characterization of Mammary Cancer Stem Cells in Mammospeheres
Eleonora Piscitelli, Cinzia Cocola, Frank Rüdiger Thaden, Paride Peluchi, Brian Gray, Giovanni Bertalot, Alberto Albertini, Rolland Reinbold, and Ileana Zucchi
19. Isolation and Culture of Primary Glioblastoma Cells from Human Tumor Specimens
Sascha Seidel, Boyan K .Garvalov, and Till Acker