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Blood, Vol. 112, Issue 12, 4503-4506, December 1, 2008
Trisomy 21 enhances human fetal erythro-megakaryocytic development
Blood Chou et al.
112: 4503
Supplemental materials for: Chou et al
Files in this Data Supplement:
- Document 1. Supplemental materials and methods (PDF, 63.5 KB)
- Table S1. Oligonucleotides used for real-time RT-PCR (PDF, 37.3 KB)
- Table S2. Hematopoietic colony assays with unfractionated mononuclear cells from control and trisomy 21 fetal livers (PDF, 38.5 KB) -
Erythroid colony-forming units (CFU-E), erythroid burst-forming units (BFU-E), granulocyte macrophage colony-forming units (CFU-GM), and megakaryocyte colony-forming units (CFU-Mk) were scored on days 8–9, 13–14, 11–14, and 11–12 respectively. Values shown are the mean colony count of triplicate assays per fetal liver (FL) specimen. GSA, gestational age; avg, average.
- Figure S1. Reconstitution of human hematopoiesis in NOD/SCID/IL-2Rγcnull mice (JPG, 115 KB)
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T21 or control fetal MNCs were transplanted into sublethally irradiated mice. At 5–10 weeks post transplantation, bone marrow chimerism for donor cell-derived lineages was determined by flow cytometric analysis for human CD45, CD33 (myeloid), CD19 (B-lymphocyte), CD41a (megakaryocyte), and glycophorin A (erythroid) surface antigen expression. CD45−/dim+ cells were analyzed for human CD41a and glycophorin A to assess human megakaryocytic and erythroid reconstitution, respectively. CD45+ cells were examined for the expression of CD19 and CD33 surface markers for human B-lymphoid and myeloid cell chimerism, respectively. Numbers indicate percentages of parent population.
- Figure S2. Colony assays of purified progenitor compartments from C (n=4) and T21 (n=4) fetal liver MNCs (JPG, 32.7 KB)
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Sorted populations (Fig. 2A) were seeded into methylcellulose colony assays to quantify BFU-E and CFU-GM progenitors. CFU-Mk were analyzed using the Megacult assay (Stem Cell Technologies). Colony count reflects colonies generated from 300 sorted progenitors/ml for BFU-E and CFU-GM cultures, and 2500 cells/ml for CFU-Mk assays. Results shown are the mean values. CMP, common myeloid progenitor, GMP, granulocyte macrophage progenitor; MEP, megakaryocyte erythroid progenitor.
- Figure S3. Gene expression analysis of selected chromosome 21-encoded hematopoietic transcriptional regulators in purified HSCs, as measured by real-time RT-PCR and normalized to levels of β-actin (JPG, 44.1 KB)
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Each bar represents an individual control (C) or trisomy 21 (T21) fetal liver sample.
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