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 populationFlow cytometry in the study of proliferation and apoptosisStem cell biology and applicationIdentification and isolation of very small embryonic-like stem cells from murine and human specimensHematopoietic stem cellsissues in enumerationHuman embryonic stem cells: long-term culture and cardiovascular differentiationLimbal stem cells and corneal regenerationFlow cytometric sorting of spermatogonial stem cellsBreast cancer stem cellsStem 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.

Awtar Krishan, Ph.D is a Professor of Pathology at the University of Miami School of Medicine. From 1965 to 1977, he was at the Sidney Farber Cancer Institute, Harvard MedicalSchool as Head of the Tissue Ultrastructure and Experimental Pathology Labs. Some of his recent work has focused on development and applications of a new flow cytometer which can measure cell volume and marker expression of stem cells in human apheresis samples. Dr. Krishan has more than 160 publications in peer reviewed journals.

H. Krishnamurthy is working as Scientific Officer 'E' and in charge of Flow Cytometry and Confocal Microscopy Facilities at the National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bangalore, India. His research interest is hormonal regulation of spermatogenesis and trafficking and signaling of gonadotropin receptors.

Satish Totey is Chief Scientific Officer, Stempeutics Research Pvt. Ltd and Director at the Manipal Institute of Regenerative Medicine inBangalore. Dr. Totey was amongst the small number of researchers who pioneered the isolation and characterization of pluripotent stem cells from human embryos. The cell lines he established have been listed in the NIH stem cell registry. He has also developed up-scaling of mesenchymal stem cells and is currently conducting phase-I and II clinical trials for myocardial infarction, leg ischemia, cerebral stroke and Multiple Sclerosis. He has published 90 research papers and holds 12US patents.

CONTRIBUTORS.

PREFACE.

1 Basics of Flow Cytometry (H. Krishnamurthy and L. Scott Cram).

2 Practical Considerations for Flow Cytometric Sorting of Stem Cells (Geoffrey W. Osborne).

3 Stem Cell Analysis and Sorting Using Side Population Analysis (William Telford).

4 Flow Cytometry in the Study of Proliferation and Apoptosis (Michael G. Ormerod and Ronald M. Hamelik).

5 Flow Cytometric Analysis of Drug Transport and Efflux in Stem Cells (Awtar Krishan and Ronald M. Hamelik).

6 Stem Cell Biology and Application (Swapnil Totey, Rajarshi Pal, Murali Krishna Mamidi, Vijayendran Govindasamy, and Satish Totey).

7 Identification and Isolation of Very Small Embryonic-like Stem Cells from Murine and Human Specimens (Ewa K. Zuba-Surma, Dong-Myung Shin, Izabella Klich, Janina Ratajczak, Magda Kucia, and Mariusz Z. Ratajczak).

8 Electronic Volume of Hematopoietic Stem Cells (Siddharth Sharma and Awtar Krishan0.

9 Hematopoietic Stem Cells: Issues in Enumeration (Michael Keeney and D. Robert Sutherland).

10 Embryonic Stem Cells: Development and Characterization (Vijay Bhaskar R. Konala, Villoo Morawala-Patell, and Aparna Khanna).

11 Human Embryonic Stem Cells: Long-Term Culture and Cardiovascular Differentiation (Maneesha Inamdar).

12 Mesenchymal Stromal Cells and Their Clinical Applications (Jyoti Kode and Vivek Tanavde).

13 Circulating Adult Stem Cells of Hematopoietic Origin for Vascular and Neural Regeneration (Lissy K. Krishnan).

14 Flow Cytometric Characterization of Neural Progenitors Derived from Human Pluripotent Stem Cells (Raj R. Rao, Sujoy K. Dhara, Shilpa Iyer, and David W. Machacek).

15 Limbal Stem Cells and Corneal Regeneration (Geeta K. Vemuganti, Murali Mohan Sagar Balla, and Shubha Tiwari).

16 Flow Cytometric Sorting of Spermatogonial Stem Cells (B. S. Srinag, J. M. Kalappurakkal, G.H. Mohan, and H. Krishnamurthy).

17 Breast Cancer Stem Cells (Devaveena Dey and Annapoorni Rangarajan).

18 Tumor Stem Cell Marker Expression in Cells from Body Cavity Fluids (Awtar Krishan, Deepti Sharma, and Ronald M. Hamelik).

INDEX.