Identification and Characterization of Cancer Stem Cells Using Flow Cytometry PDF Download

Author: Yasunari Kanda
Publisher:
ISBN:
Category : Medicine
Languages : en
Pages :

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Identification and Characterization of Cancer Stem Cells Using Flow Cytometry.

Author: Awtar Krishan
Publisher: John Wiley & Sons
ISBN: 1118148061
Category : Science
Languages : en
Pages : 317

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Book Description
A much-needed primer on the use of laser flow cytometry for stem cell analysis Laser flow cytometry is a powerful tool for rapid analysis of cells for marker expression, cell cycle position, proliferation, and apoptosis. However, no resources specifically address the use of this methodology for the study of stem cells; this is especially important as stem cell analysis involves specialized methods and staining procedures based on specific characteristics such as marker expression, cell size, drug transport, and efflux of the stem cells. Now, this book reviews these procedures, discusses the science behind them, and provides real-world examples to illustrate the usefulness of the methods. It brings together world-class experts in pathology, biophysics, immunology, and stem cell research, who draw upon their extensive experience with the methods and show examples of good data to help guide researchers in the right direction. Chapter coverage includes: Stem cell analysis and sorting using side population Flow cytometry in the study of proliferation and apoptosis Stem cell biology and application Identification and isolation of very small embryonic-like stem cells from murine and human specimens Hematopoietic stem cells—issues in enumeration Human embryonic stem cells: long-term culture and cardiovascular differentiation Limbal stem cells and corneal regeneration Flow cytometric sorting of spermatogonial stem cells Breast cancer stem cells Stem cell marker expression in cells from body cavity fluids This book is an essential resource for all graduate students, practitioners in developing countries, libraries and book repositories of universities and research institutions, and individual researchers. It is also of interest to laboratories engaged in stem cell research and use of stem cells for tissue regeneration, and to any organization dealing in stem cell and tissue regeneration research.

Author: Federica Papaccio
Publisher: Springer Nature
ISBN: 1071637304
Category :
Languages : en
Pages : 263

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Author: Kit-Man Sunny Wong
Publisher:
ISBN: 9781361302187
Category :
Languages : en
Pages :

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This dissertation, "Isolation and Characterization of Cancer Stem Cells in Non-small Cell Lung Cancer" by Kit-man, Sunny, Wong, 王傑民, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Tumor heterogeneity has long been observed and recognized as a challenge to cancer therapy. The cancer stem cell (CSC) model is one of the hypotheses proposed to explain such a phenomenon. A specific cancer stem cell marker has not been determined for non-small cell lung cancers (NSCLC), preventing the definitive evaluation of whether the biology of NSCLC is governed by the CSC model. This study aimed to analyze the expression of candidate CSC markers and using the identified putative marker, to isolate CSC and determine the applicability of the CSC model in NSCLC. The expression or activities of four putative stem cell markers, CD24, CD44, CD133 and aldehyde dehydrogenase 1 (ALDH1) were measured by flow cytometry in eight NSCLC cell lines before and after chemotherapy for 24 hours. Markers with increased expression after treatment were considered potential CSC markers and used for isolating tumor cell subpopulations from the untreated cell lines by fluorescence-activated cell sorting (FACS). Confirmatory analyses using widely acceptable methodology were performed to test for CSC properties in the marker+ and marker- subpopulations. Isolated subpopulations were further characterized by functional and phenotypic studies. Flow cytometry showed amongst the 4 markers, only ALDH1 expression was significantly enhanced by chemotherapeutic treatment, suggesting ALDH1 could be a CSC marker. Untreated ALDH1+ cells formed significantly more and larger cell spheres in non-adherent, serum-free conditions than ALDH1- cells. Likewise, higher in vitro tumorigenic ability was observed in ALDH1+ subset using colony formation assay. Furthermore, a higher resistance to cytotoxic drugs was observed in ALDH1+ compared to ALDH1- cells. In vivo studies also showed ALDH1+ cells showed higher tumorigenicity than ALDH1- cells; as few as 2,500 ALDH1+ cells formed tumor in SCID mice which were serially transplantable to 2nd and 3rd recipients, while no tumor was formed from ALDH- cells with even ten times the number of cells. Also, expression analysis revealed higher expression of the pluripotency genes, OCT4, NANOG, BMI1 and SOX9, in ALDH1+ cells. In view of previous studies reporting CD44 as a CSC marker in lung cancer, double marker selection of putative CSC was performed to compare ALDH1+CD44+ and ALDH1-CD44+ subpopulations. Results of the spheroid body formation assay and cisplatin treatment experiments revealed the ALDH1+CD44+ subpopulation possessed higher self-renewal ability and chemo-resistance. Cell migration and invasion assays showed differences between the ALDH1+CD44+ and ALDH1- CD44+ subpopulations. The significance of these observations require further investigation. In conclusion, the result showed that ALDH1 could be a marker for NSCLC stem cells as evidenced by enhanced self-renewal and differentiation abilities in ALDH1+ subpopulation. Furthermore, this study observed the presence of at least two potential stem cell subpopulations in NSCLC cells with differential selfrenewal, chemotherapy resistance and cell mobility properties. Further investigations are required to validate th

Author: Susan Friedman
Publisher:
ISBN:
Category :
Languages : en
Pages : 46

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Author: Sharmila A. Bapat
Publisher: John Wiley & Sons
ISBN: 0470391561
Category : Science
Languages : en
Pages : 274

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Book Description
Because the concept and discoveries of cancer stem cells are relatively new, scientists and researchers need an introduction to this dynamic area. Cancer Stem Cells presents a consolidated account of the research done to date and recent progresses in the studies of cancer stem cells. Such a presentation facilitates a better understanding of and draws attention to stem cell and cancer biology - two fields that enhance, move, and evolve into each other continuously. It provides an informative study in designing approaches to apply stem cell principles to cancer biology while offering an overview of the challenges in developing combination stem and cancer biology targets for therapeutics. This book serves as a primer for new researchers in the field of cancer biology.

Author: Virginia Litwin
Publisher: John Wiley & Sons
ISBN: 0470922788
Category : Medical
Languages : en
Pages : 404

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This book covers the unique application of flow cytometry in drug discovery and development. The first section includes two introductory chapters, one on flow cytometry and one on biomarkers, as well as a chapter on recent advances in flow cytometry. The second section focuses on the unique challenges and added benefits associated with the use of flow cytometry in the drug development process. The third section contains a single chapter presenting an in depth discussion of validation considerations and regulatory compliance issues associated with drug development.

Author: Ryan Jackson Ward
Publisher:
ISBN: 9780494977903
Category :
Languages : en
Pages :

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Author: Wing-Yuen Ho
Publisher: Open Dissertation Press
ISBN: 9781361007433
Category : Medical
Languages : en
Pages : 258

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This dissertation, "Identification and Characterization of CD90⁺ Cancer Stem Cells in Hepatocellular Carcinoma" by Wing-yuen, Ho, 何永源, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Hepatocellular carcinoma (HCC) is one of the most devastating malignancies worldwide with increasing incidences in both developed and developing countries. Survival rates have not been significantly improved over the past decades in spite of advances in detection and therapies for this disease, suggesting that current treatments may target the wrong cells, and miss the cancer stem cells (CSCs). The cancer stem cell hypothesis presents that tumor formation, proliferation and propagation are driven by a rare subpopulation of chemoresistant CSCs that are not killed by conventional therapies and go on to cause disease relapse. The objective of this study was to identify and characterize CSCs in HCC cell lines and human liver tumor specimens using CD90 as a potential marker. The number of CD90+ cells present in HCC cell lines was found to positively correlate with tumorigenicity potentials. Injection of as few as 2,000 sorted CD90+ cells from HCC cell lines resulted in the formation of tumor nodules in nude mice, whereas no tumors formed for CD90ˉcells in the same model. The tumor xenograft generated by injection of CD90+ cells sorted from previous xenograft in a serial xenotransplantation assay exhibited recapitulation of tumor heterogeneity to original primary tumor and consistent proportion of CD90+ and CD90ˉ cells which demonstrated self-renewal and differentiation capacities of CD90+CSCs. CD45ˉCD90+ cells were detected (0.03%-6.2%) in human liver tumor specimens, but were only present in minute quantities in normal, cirrhotic and non-tumorous tissues. More importantly, CD45ˉCD90+ cells sorted from primary HCC tumor also displayed tumorigenicity, self-renewal and lineage differentiation capacities. CD90+CSCs were found to be more resistant to therapeutic drugs compared to CD90- cells, as reflected by the results of enrichment of the CD90+ CSCs and longer survival rates after chemotherapeutic treatment. The high expression of genes, such as OCT4, MRP3, ABCG2, AKT1, BirC5, BCL2, HA and CD44, in CD90+CSCs may mediate chemoresistance. The majority of CD90+ cells co-expressed CD44, another stem cell marker. Blocking CD44 activities by anti-CD44 antibody increased apoptosis of CD90+ CSCs, sensitized CD90+CSCs to chemotherapeutic drugs in vitro, and decreased tumorigenic and metastatic potentials of CD90+CSCs in vivo, indicating that a therapeutic potential of targeting CD44. However side effects may be problematic due to the endogenous expression of CD44 in healthy tissues and normal lymphocytes. To identify novel gene targets specific to liver CSCs, a sensitive RNA-sequencing (RNA-Seq) technique was used to compare the gene expression profiles between CD90+CSCs sorted from HCC primary tumors and CD90+cells from adjacent non-tumorous tissue (CD90+NTSCs). The up-regulated genes in CD90+CSCs were associated with lipid metabolism, inflammation, and drug resistance. Among the differentially expressed genes, glypican-3 (GPC3) was specifically elevated in CD90+CSCs but not in CD90+NTSCs. Therefore, GPC3 could be a promising gene candidate for HCC therapy as targeting GPC3 should not induce damage to normal liver stem cells. In summary, CD90 is a liver CSCs marker. Identification of CD90+ CSCs in HCC provides new insight into cellular basis of hepatocarcinogenesis, recurrence and metastasis, which opens new avenues for the design of future CSC-targeted therapies. D

Author: Mohan C. Vemuri
Publisher: Springer Science & Business Media
ISBN: 1588297446
Category : Medical
Languages : en
Pages : 815

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The scope for improving health care using stem cell therapies is thrilling, but has considerable technical challenges and methodological constraints that need to be addressed. Keeping with the tradition of Humana Press to bring these developments to the forefront in a timely manner, this book presents scientific advances in stem cell methods for a wider use by novice and expert scientists, through the series of Methods in Molecular Biology.