New PDF release: Hematopoietic Stem Cell Protocols

By Klug C.A., Jordan C. (ed.)

Univ. of Alabama, Birmingham. deals a wide selection of step-of-step tools for learning hematopoietic stem cells. info equipment for the purification and genetic amendment of stem cells and contains stem mobile enlargement protocols. DNLM: Hematopoietic Stem Cells - cytology.

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I. Incubate 20 min at 37°C. If clumps are sill present, repeat pipetting procedure. j. Filter through a coarse (70–100 µ) filter, adding D-PBS as necessary. k. Spin, remove supernatant, and resuspend in D-PBS with FBS for ficolling. Cord blood: Cord blood samples are especially difficult to process, as the density of the cells can vary over a wide range. Cord blood may also have very high platelet counts which lead to clumping, despite the presence of anticoagulants. Cells should be suspended in 10% anticoagulant citrate dextrose (ACD) or 5% ACD plus 1% Human Albumin in saline.

Proc. Natl. Acad. Sci. USA 87, 3584–3588. 6. Terstappen, L. W. M. , Lansdorp, P. , and Loken, M. R. (1991) Sequential generations of hematopoietic colonies derived from single nonlineage-committed CD34+CD38– progenitor cells. Blood 77, 1218–1227. 7. , Eaves, C. , Sutherland, H. , and Lansdorp, P. M. (1991) Separation of functionally distinct subpopulations of primitive human hematopoietic cells using rhodamine-123. Exp. Hematol. 19, 338–342. 8. Lansdorp, P. M. and Dragowska W. (1992) Long-term erythropoiesis from constant numbers of CD34+ cells in serum-free cultures initiated with highly purified progenitor cells from human bone marrow.

22. 23. 24. 25. 26. Thomas et al. recombinant human granulocyte-macrophage colony-stimulating factor. Blood 74, 1905–1914. Yin, A. , Leary ,A. , et al. (1997) AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood 90, 5002–5012. Thomas, T. , Fairhurst, M. , and Lansdorp, P. M. (1997) Rapid single step immunomagnetic isolation of highly enriched primitive human hematopoietic progenitors. Blood 9, 347b (Abstract). Hogge, D. , Lansdorp, P. , and Eaves, C. J. (1996) Enhanced detection, maintenance, and differentiation of primitive human hematopoietic cells in cultures containing murine fibroblasts engineered to produce human steel factor, interleukin-3, and granulocyte colony-stimulating factor.

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Hematopoietic Stem Cell Protocols by Klug C.A., Jordan C. (ed.)

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