This volume on stepwise techniques to culture adult human stem cells begins with the basic methods to culture adult stem cells followed by related methods to supplement stem cell culture. This and future volumes of techniques will allow laboratories across scientific entities – academic and commercial – to adapt the described methods for similar methods with cells from other species. The standardized techniques will allow across the board interpretation of other technologies such as Omics studies. These methods can be a starting platform for culturing stem cells even in laboratories that might not be familiar with the techniques.
Scientific and early clinical trials strongly favor the development of translational studies with adult stem cells. A major impediment to progress in the field of regenerative medicine is a lack of standardized methods to culture specific stem cells. Such documented methods would allow for scientific rigor and reproducibility to enhance efficient and safe translation of stem cells to patients. This book fills that gap.
Introduction: Core Techniques in Stem Cell Culture
Pranela Rameshwar
1. Isolation of Hematopoietic Stem Cells
Bobak Shadpoor, Lauren S. Sherman, Sebastian Acevedo, Yannick Kenfack, and Pranela Rameshwar
2. Isolation Protocol for CD133+ and CD34+ Very Small Embryonic Like Cells (VSEL) from Human Umbilical Cord Blood
Kamila Bujko, Vira Chumak, Mariusz Z. Ratajczak, and Magdalena Kucia
3. Isolation and Expansion of Mesenchymal Stem Cells from the Adult Bone Marrow Aspirate
Lauren S. Sherman, Harrison C. Rosemond, andPranela Rameshwar
4. Manual Isolation and Expansion of Adipose Derived Stem Cells from Adipose Tissue
Lauren S. Sherman, Derek J. Woloszyn, Pranela Rameshwar, and Edward S. Lee
5. Isolating Compact Bone-derived Mesenchymal Stem Cells from Rodent and Rabbit Femurs
Najerie McMillian, Joseph S. Fernandez-Moure, and Taylor Hudson
6. Isolating Dental Pulp Stem Cells
Ioanna Tsolaki, Darling Rojas, Adam Eljarrah, and Pranela Rameshwar
7. A Murine System for Somatic Cellular Reprogramming into Induced Pluripotent Stem Cells
Dahui Wang, Edward A. Gonzalez, and Jean-Pierre Etchegaray
8. Isolation and Expansion Of Placenta Stem Cells (P-MSCs)
B. Shadpoor, L. S. Sherman, M. P. Romagano, Shauna F. Williams, K. Krishnamoorthy, K. Powell, Y. K. Kenfack, and P. Rameshwar
9. Expanding Human Mesenchymal Stem Cells from the Umbilical Cord
Lauren S. Sherman, Andrew Petryna, Matthew P. Romagano, and Pranela Rameshwar
10. Laboratory Isolation of Leukemia Cancer Stem Cell
Michael Haddadin and Shyam A. Patel
11. Isolation and Characterization of Cancer Stem cells from Glioblastoma Multiforme and Breast Cancer Cell Lines
Vibha Harindra Savanur, Alejandra I. Ferrer, andPranela Rameshwar
12. Bone Marrow Aspirate for Isolation of Mesenchymal Stem Cells
Lauren S. Sherman, Andrew Petryna, and Joshua Kra
13. Culturing Human Bone Marrow Stromal Cells
Yannick Kenfack, Lauren S. Sherman, Bobak Shadpoor,Andrew Petryna, Sami Souyah, Ella Einstein, Stephanie Perricho, and Pranela Rameshwar
14. Isolating Mononuclear Cells by Ficoll-Hypaque Density Gradient
Andrew Petryna, Lauren S. Sherman, Anushka Sarkar,Bobak Shadpoor, Yannick Kenfack, and Pranela Rameshwar
15. Phenotypic and Multipotent Characterization of Bone Marrow-derived Mesenchymal Stem Cells
Anuska Sarkar, Andrew Petryna, Bobak Shadpoor, Lauren S. Sherman, Pranela Rameshwar
16. Cryopreservation of Relatively Small Number of Stem Cells
Lauren S. Sherman and Pranela Rameshwar
17. Cryopreservation of Mobilized Peripheral Blood Stem Cells
Seda Ayer, Khadidiatou Guiro, Steven S. Greco, Oleta A. Sandiford, Lauren S. Sherman, Garima Sinha, Nykia D. Walker, and Pranela Rameshwar
18. Manual Cell Count
Andrew Petryna, Lauren S. Sherman, and Pranela Rameshwar
Biography
Pranela Rameshwar is a tenured professor of medicine, Division of Hematology and Oncology at Rutgers New Jersey Medical School, Rutgers. She received her B.Sc. degree in medical microbiology from the University of Wisconsin at Madison and Ph.D. in biology from Rutgers University, New Jersey. She performed postdoctoral studies in hematopoiesis at New Jersey Medical School. Dr. Rameshwar’s research interest in stem cell biology began with her doctoral thesis and continues with research on breast cancer dormancy, neural regulation of hematopoiesis, and clinical application of adult human mesenchymal stem cells. Her research continues to be funded by federal, state, and other agencies. Dr. Rameshwar has edited books and authored >200 publications, which include original articles, reviews, editorials and book chapters. Dr. Rameshwar initiated a certificate program in stem cell biology and directs four graduate level courses in stem cell biology.
