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Bone Marrow and Stem Stell Transplantation (Methods in by Meral Beksac

By Meral Beksac

This quantity is a compendium of state-of-the-art molecular equipment for the winning transplantation of hematopoietic stem cells. The participants are world-renown leaders within the box. They describe promising instruments for stem telephone transplant learn versions, similar to in vivo bioluminescence imaging. They talk about HLA typing, PCR-SSP typing, and HLA antigens. This quantity is a useful resource for biochemists, molecular biologists, and clinicians.

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189, 51–363. Sipkins, D. , Wu, J. , et al. (2005) In vivo imaging of specialized bone marrow endothelial microdomains for tumour engraftment. Nature 435, 969– 973. Sorg, B. , Moeller, B. , and Dewhirst, M. W. (2005) Hyperspectral imaging of hemoglobin saturation in tumor microvasculature and tumor hypoxia development. J. Biomed. Opt. 10, 44,004. , Brotz, T. , and Germain, R. N. (2002) Dynamic imaging of T cell-dendritic cell interactions in lymph nodes. Science 296, 1873–1876. , Wahl, R. , et al.

13. Spin at 1200 rpm (approx 310g) for 5 min at 4°C. 14. 5 ϫ 106 cells/mL (see Note 6). 15. 5 ϫ 106 cells/well in a 24-well plate, 1 mL per well. Incubate at 37°C/ 5% CO2 overnight. DAY 6: 16. Remove the supernatant from the transfected packaging cell line. Replace with fresh media and return the cells to the incubator. 17. To the supernatant, add polybrene to 5 µg/mL and HEPES to 10 mM. Mix thoroughly. 18. 45-µm filter. Imaging Cell Fates and Function 29 19. Centrifuge the bone marrow cells at 2100 rpm (approx 950g) for 5 min.

An alternative to using a stably transfected “viral packaging cell line” is to use a second plasmid that contains only the genes encoding envelope and packaging proteins. ) In the cells that receive both plasmids, the presence of ψ-flagged DNA along with envelope and packaging proteins leads to the generation of viral particles. As with the stably transfected packaging line, the fact that the genes encoding envelope and packaging proteins are not present on the ψ-flagged DNA means that those genes will not be included in the viral particles and will not be present in the target cell.

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