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Effects of anoxia on megakaryocyte progenitors derived from cord blood CD34pos cells

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Saxonhouse MA, Rimsza LM, Christensen RD, Hutson AD, Stegner J, Koenig JM, Sola MC. Effects of anoxia on megakaryocyte progenitors derived from cord blood CD34pos cells.

Eur J Haematol 2003: 71: 359–365. © Blackwell Munksgaard 2003. Abstract: Background:

Severe hypoxic insults to the fetus and neonate are associated with the development of thrombocytopenia. The thrombocytopenia in some cases is the result of disseminated intravascular coagulation, but that mechanism fails to account for all, perhaps the majority, of cases. Objective:

We hypothesized that human fetal megakaryocyte (Mk) progenitors are directly adversely affected by transient anoxia. Design and methods:

To test this, we isolated CD34pos cells from the umbilical cord blood of 10 healthy term neonates, and exposed these to 0% or 20% O2 for 24 h, with or without recombinant thrombopoietin (rTpo, 50 ng/mL). After 24 h, a portion of the CD34pos cells were harvested for flow cytometric evaluation of apoptosis. The remaining cells were cultured for an additional 10–12 days, under normoxic conditions, in a collagen-based serum-free system containing rTpo, IL-3, and IL-6. In this way, we sought to determine the effect of transient anoxia on clonogenic capacity of Mk progenitors. Results:

Contrary to our hypothesis, anoxia did not increase either apoptosis or cell death of the CD34pos cells. The addition of rTpo was protective, with a significant decrease in apoptosis and cell death (P < 0.0001), and an increase in the number of Mk colonies cultured (P = 0.04). There was no difference between the normoxic and anoxic groups in proliferative potential of the Mk progenitor cells. Conclusions:

The thrombocytopenia observed in neonates following an acute hypoxic event is not likely due to a direct deleterious effect of hypoxia on Mk progenitors.
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Keywords: CD34pos cells; anoxia; apoptosis; megakaryocyte progenitors; rTpo

Document Type: Research Article

Affiliations: 1: Department of Pediatrics, University of Florida, Gainesville, FL 2: Department of Pathology, University of Arizona, Tucson, AZ 3: Department of Pediatrics, University of South Florida and All Children's Hospital, St Petersburg, FL 4: Division of Biostatistics, University at Buffalo, Buffalo, NY, USA

Publication date: November 1, 2003

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