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Blood, Vol. 105, Issue 1, 85-94, January 1, 2005
Different steroids co-regulate long-term expansion versus terminal differentiation in primary human erythroid progenitors
Blood Leberbauer et al.
105: 85
Supplemental materials for: Leberbauer et al, Vol 105, Issue 1, 85-94
Files in this Data Supplement:
- Table S1. Progression of cell surface marker expression in developing erythroblast cultures (PDF, 59 KB) -
Aliquots of proliferating cells were harvested at the times indicated, stained with combinations of fluorescently labeled antibodies against markers characteristic for different hematopoietic lineages and stages of development, and subjected to flow cytometry. Markers are characteristic for CD56, natural killer cells; CD8, cytotoxic T cells; CD4, T-helper cells; CD45RA, mixed-lineage, nonerythroid (in combination with CD19, B-cell specific); CD116, monocytic progenitors; and CD15, granulocytes.
- Figure S1. Long-term expansion of human erythroid progenitors under standard proliferation conditions (JPG, 13 KB)
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Cell size and proliferation kinetics in the presence or absence of cholesterol-rich lipids were determined by daily measurements in an electronic cell counter. Cumulative cell numbers are calculated as described in "Materials and methods"; error bars are SD of mean, n|=|5. Cultures without lipid were terminated at day 30 due to excessive cell death.
- Figure S2. Proliferation of male versus female human erythroblasts on addition of androgen (JPG, 12 KB)
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Cell numbers from cultures of male versus female cord blood donors (pregrown for 8 days in the absence of sex steroids) were counted daily in an electronic cell counter, and cumulative cell numbers were determined (Figure 2 and "Materials and methods"). Cells were cultivated either under standard proliferation conditions (control, diamonds) or in the presence of 1 µM testosterone (squares) or testosterone plus the androgen antagonist Cpa (1 µM; circles); error bars are SD of mean, n|=|3.
- Figure S3. Reduced proliferation capacity of human erythroblasts in the presence of serum (JPG, 9 KB)
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Erythroblasts were cultivated after addition of 3% human serum (HS) or, as a control, under standard proliferation conditions (Table 1). Cell numbers to determine proliferation kinetics (A) and cell volumes (B) were monitored daily, using an electronic cell counter.
- Figure S4. Enhanced spontaneous differentiation of human erythroblasts in the presence of T3 (JPG, 9 KB)
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Erythroblasts were cultivated after addition of T3 or, as a control, under standard proliferation conditions (Table 1). Cell numbers to determine proliferation kinetics (A) and cell volumes (B) were monitored daily, using an electronic cell counter.
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