Molecular Biology of the Cell, Fourth Edition Tom Clancy's Splinter Cell Outsmarting the Midlife Fat Cell : Winning Weight Control Strategies for Women Over 35 to Stay Fit Through Menopause Outsmarting the Female Fat Cell : The First Weight-Control Program Designed Specifically for Women Molecular Cell Biology, Fifth Edition The Lives of a Cell: Notes of a Biology Watcher The Jerk with the Cell Phone: A Survival Guide for the Rest of Us The Human Embryonic Stem Cell Debate: Science, Ethics, and Public Policy (Basic Bioethics) The Prisoner in the Third Cell

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• Stem Cells 2003;21:272-280 www. ...
• Key Words: Embryonic stem cells · Hemangioblast · Thrombopoietin · c-Mpl.
• Because thrombopoietin (TPO) and its receptor, c-Mpl, regulate primitive hematopoietic populations, including bone marrow hematopoietic stem cells, we investigated whether TPO acts on the hemangioblasts that derive from differentiation of embryonic stem cells in vitro. ... When replated under the appropriate conditions, TPO-stimulated BL-CFC gave rise to secondary hematopoietic colonies, as well as endothelial cells, confirming their nature as hemangioblasts. ...
• Although it was initially thought to be a lineage-dominant factor primarily affecting megakaryopoiesis 1–3 , numerous in vitro studies have established that TPO plays an important role in the maintenance and expansion of hematopoietic stem cells (HSCs) 4–10 . ... demonstrated that c-mpl-/- mice were severely deficient in spleen colony-forming cells (CFC-S) as well as long-term repopulating HSCs, and that engraftment ability was extinguished on serial transfer into secondary and tertiary hosts, suggesting that the absence of c-Mpl signaling might affect the self-renewal properties of these cells 15 . ...
• In normal mice, c-Mpl is expressed in megakaryocytes, platelets, and primitive hematopoietic cells of fetal liver and BM 16, 17 . ... Interestingly, low levels of expression have also been detected in endothelial cells derived from the umbilical cord 16, 19 and more abundantly in liver endothelial cells 20 . ...
• The close association between endothelial cells and primitive hematopoietic precursors in the embryonic yolk sac suggests that the endothelium plays a critical role in early hematopoietic development, providing the microenvironment required for stem cell proliferation and differentiation. ... These observations have led to the hypothesis that a common progenitor, the hemangioblast, gives rise to both endothelial and hematopoietic cells 26, 27 . Recently, a cell with properties of the hemangioblast has been identified during in vitro differentiation of embryonic stem (ES) cells into embryoid bodies (EBs) 28 . This precursor, referred to as the blast colony-forming cell (BL-CFC) forms in response to vascular endothelial growth factor (VEGF) and stem cell factor (SCF), and represents a transient population of cells that stand at the juncture of the endothelial and hematopoietic lineages 29 . ...
• In this study, we demonstrated that TPO and its receptor are expressed at the yolk sac stage of hematopoiesis in the embryo, and that TPO stimulates hemangioblast formation during differentiation of ES cells in vitro. ...
• Growth and Differentiation of ES Cells.
• For differentiation cultures, the cells were dissociated with trypsin (0. ... Cells were washed three times with phosphate-buffered saline (PBS) and EBs were generated by plating 104 cells/ml in Iscove’s modified Dulbecco’s medium (IMDM; Sigma) with 15% fetal calf serum (FCS; Sigma), 50 µg/ml ascorbic acid (Sigma), 200 µg/ml iron-saturated transferrin (Sigma), 4. ...

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