Methods in Stem Cell Biology - Part B

1st Edition - August 9, 2022

Editors: Ilio Vitale, Gwenola Manic, Lorenzo Galluzzi

Hardback ISBN:

9 7 8 - 0 - 3 2 3 - 9 0 0 1 8 - 8

eBook ISBN:

9 7 8 - 0 - 3 2 3 - 9 0 0 1 9 - 5

Methods in Cancer Stem Cell Biology: Part B, Volume 171 in the Methods in Cell Biology series highlights advances in the field, with this new volume presenting interesting… Read more

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Methods in Cancer Stem Cell Biology: Part B, Volume 171 in the Methods in Cell Biology series highlights advances in the field, with this new volume presenting interesting chapters on timely topics, including Orthotopic brain tumor models derived from glioblastoma stem-like cells, RNA sequencing in hematopoietic stem cells, Generation of inducible pluripotent stem cells from human dermal fibroblasts, In vitro preparation of dental pulp stem cell grafts combined with biocompatible scaffolds for tissue engineering, Gene expression knockdown in chronic myeloid leukemia stem cells, Identification and isolation of slow-cycling GSCs, Assessment of CD133, EpCAM, and much more.

Cover image
Title page
Table of Contents
Copyright
Contributors
Chapter 1: Multiple gene knockdown strategies for investigating the properties of human leukemia stem cells and exploring new therapies
Abstract
1: Introduction
2: Materials
3: Methods
4: Notes
5: Concluding remarks
Acknowledgments
Disclosures
References
Chapter 2: Evaluation of cancer stem cells markers expression in HCC trough real-time polymerase chain reaction
Abstract
1: Introduction and rationale
2: Materials
3: Methods
4: Results and discussion
Disclosures
References
Chapter 3: Reverse Phase Protein Arrays in cancer stem cells
Abstract
1: Introduction
2: Materials
3: Methods
4: Notes
5: Concluding remarks
Disclosures
References
Chapter 4: Isolation of cancer stem cells from skin squamous cell carcinoma
Abstract
1: Introduction
2: Materials
3: Methods
4: CSCs assays
5: Precautions to be taken during CSC isolation to avoid commonly faced problems
6: Concluding remarks
Disclosures
References
Chapter 5: Immunoblotting-assisted assessment of JAK/STAT and PI3K/Akt/mTOR signaling in myeloproliferative neoplasms CD34+ stem cells
Abstract
1: Introduction
2: Materials
3: Isolation of mononuclear cells (MNCs) from human peripheral blood by density gradient centrifugation
4: Immunomagnetic separation of hematopoietic stem cells (CD34 +) from MNCs
5: Protein extraction and quantification from CD34 + cells
6: Sample preparation and SDS-PAGE Western blotting
7: Membranes probing to detect phosphorylated proteins of JAK/STAT and PI3K/mTOR/Akt pathway
8: Phosphorylated protein revelation and data acquisition
9: Membrane stripping and re-probing to detect total protein of JAK/STAT and PI3K/mTOR/Akt pathway
10: Human colony forming unit (CFU) assay of hematopoietic stem cells (CD34 +)
11: Notes
12: Concluding remarks
Disclosures
References
Chapter 6: Isolation of cancer stem cells from cultured breast cancer cells and xenografted breast tumors based on aldehyde dehydrogenase activity
Abstract
1: Background
2: The AldefluorTM assay to isolate putative cancer stem cells
3: The importance of ALDH1A1 and ALDH1A3 in breast cancer
4: Conclusions
Acknowledgments
References
Chapter 7: Intracerebroventricular transplantation of human iPSC-derived neural stem cells (hiPSC-NSCs) into neonatal mice
Abstract
1: Introduction
2: Materials
3: Methods
4: Notes
5: Concluding remarks
Acknowledgments
References
Chapter 8: Quantifying epithelial-mesenchymal heterogeneity and EMT scoring in tumor samples via tyramide signal amplification (TSA)
Abstract
1: Introduction
2: Materials
3: Methods
4: Imaging and analysis
5: Quantification
6: Concluding Remarks
7: Limitations
8: Notes
References
Chapter 9: Immunofluorescence staining of colorectal cancer patient-derived organoids
Abstract
1: Introduction
2: Materials
3: Method
4: Notes
5: Concluding remarks
Disclosures
References
Chapter 10: Isolation of murine bone marrow hematopoietic stem and progenitor cell populations via flow cytometry
Abstract
1: Introduction
2: Materials
3: Methods
4: Notes
5: Concluding remarks
Acknowledgments
Conflict of interest
Disclosures
References
Chapter 11: Limiting dilution assay to quantify the self-renewal potential of cancer stem cells in hepatocellular carcinoma
Abstract
1: Introduction
2: Materials
3: Method
4: NOTES
Funding
References
Chapter 12: Methods to isolate adipose tissue-derived stem cells
Abstract
1: Introduction
2: Stromal vascular fraction (SVF) and adipose tissue-derived stem cells (ADSCs)
3: Enzymatic isolation of ADSCs: An overview
4: Non-enzymatic isolation of SVF and ADSCs
5: Novel non-enzymatic systems of ADSCs isolation
6: Conclusions
References
Chapter 13: Generation of neuronal/glial mixed cultures from human induced pluripotent stem cells (hiPSCs)
Abstract
1: Introduction
2: Materials
3: Methods
4: Expected results
5: Notes
6: Concluding remarks
Acknowledgments
References

Ilio Vitale

Dr Ilio Vitale is an esteemed oncologist. He is a member of the University of Rome and associated to the Regina Elena National Cancer Institute based in Italy.

Affiliations and expertise

Regina Elena National Cancer Institute, Rome and Department of Biology, University of Rome, Italy

Gwenola Manic

Gwenola Manic received her Ph.D. from the University of Paris Sud/Paris XI (France) in 2012 studying the impact of the design of HIV-1-derived vectors and DNA repair machinery on lentiviral expression. At present she is working as a postdoctoral fellow at the Department of Biology at the Università degli Studi di Roma “Tor Vergata” in Rome (Italy) on the contribution of chromosomal instability in tumorigenesis and the therapeutic targeting of cancer stem cells.

Affiliations and expertise

Candiolo Cancer Institute, FPO -IRCCs, Candiolo, Italy

Lorenzo Galluzzi

Lorenzo Galluzzi is Assistant Professor of Cell Biology in Radiation Oncology at the Department of Radiation Oncology of the Weill Cornell Medical College, Honorary Assistant Professor Adjunct with the Department of Dermatology of the Yale School of Medicine, Honorary Associate Professor with the Faculty of Medicine of the University of Paris, and Faculty Member with the Graduate School of Biomedical Sciences and Biotechnology of the University of Ferrara, the Graduate School of Pharmacological Sciences of the University of Padova, and the Graduate School of Network Oncology and Precision Medicine of the University of Rome “La Sapienza”. Moreover, he is Associate Director of the European Academy for Tumor Immunology and Founding Member of the European Research Institute for Integrated Cellular Pathology. Galluzzi is best known for major experimental and conceptual contributions to the fields of cell death, autophagy, tumor metabolism and tumor immunology. He has published over 450 articles in international peer-reviewed journals and is the Editor-in-Chief of four journals: OncoImmunology (which he co-founded in 2011), International Review of Cell and Molecular Biology, Methods in Cell biology, and Molecular and Cellular Oncology (which he co-founded in 2013). Additionally, he serves as Founding Editor for Microbial Cell and Cell Stress, and Associate Editor for Cell Death and Disease, Pharmacological Research and iScience.

Affiliations and expertise

Assistant Professor of Cell Biology in Radiation Oncology, Department of Radiation Oncology, Weill Cornell Medical College, NY, USA