By Awtar Krishan
A much-needed primer at the use of laser movement cytometry for stem telephone analysis
Laser move cytometry is a strong device for swift research of cells for marker expression, cellphone cycle place, proliferation, and apoptosis. even though, no assets particularly tackle using this system for the research of stem cells; this can be particularly very important as stem phone research consists of really expert tools and marking systems in keeping with particular features corresponding to marker expression, mobilephone measurement, drug delivery, and efflux of the stem cells.
Now, this ebook experiences those tactics, discusses the technological know-how at the back of them, and offers real-world examples to demonstrate the usefulness of the equipment. It brings jointly world-class specialists in pathology, biophysics, immunology, and stem telephone learn, who draw upon their large event with the equipment and express examples of fine information to assist advisor researchers within the correct path.
bankruptcy insurance contains:
- Stem mobilephone research and sorting utilizing aspect inhabitants
- Flow cytometry within the examine of proliferation and apoptosis
- Stem cellphone biology and alertness
- 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 tradition and cardiovascular differentiation
- Limbal stem cells and corneal regeneration
- Flow cytometric sorting of spermatogonial stem cells
- Breast melanoma stem cells
- Stem mobilephone marker expression in cells from physique hollow space fluids
This e-book is a vital source for all graduate scholars, practitioners in constructing nations, libraries and publication repositories of universities and examine associations, and person researchers. it's also of curiosity to laboratories engaged in stem phone examine and use of stem cells for tissue regeneration, and to any association dealing in stem phone and tissue regeneration research.Content:
Chapter 1 fundamentals of movement Cytometry (pages 1–12): H. Krishnamurthy and L. Scott Cram
Chapter 2 useful concerns for movement Cytometric Sorting of Stem Cells (pages 13–23): Geoffrey W. Osborne
Chapter three Stem phone research and Sorting utilizing aspect inhabitants research (pages 25–43): William Telford
Chapter four movement Cytometry within the examine of Proliferation and Apoptosis (pages 45–60): Michael G. Ormerod and Ronald M. Hamelik
Chapter five movement Cytometric research of Drug delivery and Efflux in Stem Cells (pages 61–74): Awtar Krishan and Ronald M. Hamelik
Chapter 6 Stem telephone Biology and alertness (pages 75–89): Swapnil Totey, Rajarshi friend, Murali Krishna Mamidi, Vijayendran Govindasamy and Satish Totey
Chapter 7 id and Isolation of Very SmaU Embryonic?like Stem Cells from Murine and Human Specimens (pages 91–101): Ewa okay. Zuba?Surma, Dong?Myung Shin, Habella Klich, Janina Ratajczak, Magda Kucia and Mariusz Z. Ratajczak
Chapter eight digital quantity of Hematopoietic Stem Cells (pages 103–114): Siddharth Sharma and Awtar Krishan
Chapter nine Hematopoietic Stem Cells: matters in Enumeration (pages 115–134): Michael Keeney and D. Robert Sutherland
Chapter 10 Embryonic Stem Cells: improvement and Characterization (pages 135–159): Vijay Bhaskar R. Konala, Villoo Morawala?Patell and Aparna Khanna
Chapter eleven Human Embryonic Stem Cells: Long?Term tradition and Cardiovascular Differentiation (pages 161–173): Maneesha Inamdar
Chapter 12 Mesenchymal Stromal Cells and Their scientific functions (pages 175–188): Jyoti Kode and Vivek Tanavde
Chapter thirteen Circulating grownup Stem Cells of Hematopoietic foundation for Vascular and Neural Regeneration (pages 189–209): Lissy okay. Krishnan
Chapter 14 move Cytometric Characterization of Neural Progenitors Derived from Human Pluripotent Stem Cells (pages 211–222): Raj R. Rao, Sujoy ok. Dhara, Shilpa Iyer and David W. Machacek
Chapter 15 Limbal Stem Cells and Corneal Regeneration (pages 223–240): Geeta okay. Vemuganti, Murali Mohan Sagar Baila and Shubha Tiwari
Chapter sixteen stream Cytometric Sorting of Spermatogonial Stem Cells (pages 241–250): B. S. Srinag, J. M. Kalappurakkal, G.H. Mohan and H. Krishnamurthy
Chapter 17 Breast melanoma Stem Cells (pages 251–270): Devaveena Dey and Annapoorni Rangarajan
Chapter 18 Tumor Stem telephone Marker Expression in Cells from physique hollow space Fluids (pages 271–277): Awtar Krishan, Deepti Sharma and Ronald M. Hamelik
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Additional resources for Applications of Flow Cytometry in Stem Cell Research and Tissue Regeneration
Add inhibitor (verapamil or fumitremorgin C) to the control sample. At the concentrations given above, this is a 1 : 200 dilution for both inhibitors (5 \iLlmL). Incubate at 37°C for 15 minutes. 4. Add Hoechst 33342 or DCV to both samples. For Hoechst 33342, this is 5 μL· of a 1-mg/mL stock solution per 1 mL of cells. For DCV, this is 2 [iL of a 5 mM stock solution per 1 mL of cells. Incubate for 90 minutes at 37°C with periodic shaking. 5. Transfer the tubes to a centrifuge and spin at 400 x RCF (relative centrifugal force) for 7 minutes to pellet the cells.
Go to the Instrument menu, then Instrument Setup, then the Create Compensation controls. The Create Compensation controls will appear, showing all of the parameters chosen (including Hoechst/DCV blue and red) (Figure 7C). 6. Delete the Hoechst/DCV blue and red parameters from the Create Compensation Controls panel. They will still be present on the Instrument Parameters panel. Press OK (Figure 7D). 7. Run the unlabeled and single color control samples as usual, with no Hoechst/DCV single color controls (although the single controls must also be Hoechst 33342 or DCV positive).
Characterization of a side population of cancer cells from human gastrointestinal system. Stem Cells 24:506-513. Haraguchi N, Inoue H, Tanaka F, Mimori K, Utsunomiya T, Sasaki A, Mori M. 2006b. Cancer stem cells in human gastrointestinal cancers. Hum Cell 19:24-29. REFERENCES 41 Ho MM, Ng AV, Lam S, Hung JY. 2007. Side population in human lung cancer cell lines and tumors is enriched with stem-like cancer cells. Cancer Res 67:4827-4833. Hoshi N, Kusakabe T, Taylor BJ, Kimura S. 2007. Side population cells in the mouse thyroid exhibit stem/progenitor cell-like characteristics.
Applications of Flow Cytometry in Stem Cell Research and Tissue Regeneration by Awtar Krishan