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Blood, Vol. 109, Issue 2, 693-702, January 15, 2007
Human mesenchymal stem cells isolated from bone marrow and lymphoid organs support tumor B-cell growth: role of stromal cells in follicular lymphoma pathogenesis
Blood Amé-Thomas et al.
109: 693
Supplemental materials for: Amá-Thomas et al, Vol 109, Issue 2, 693-702
Files in this Data Supplement:
- Table S1. Proliferation of primary purified FL B cells cocultured with stromal cells (PDF, 53.6 KB) -
Purified FL B cells (1.5 × 105 cells/well) were cultured in RPMI1640/10% FCS alone or cocultured with stromal cells pretreated or not with TNF/LT. After 4 days of culture, cells were pulsed with 1 µCi/well (0.037 MBq) tritiated thymidine (3H-TdR). 3H-TdR incorporation of stromal cells alone and 3H-TdR incorporation of FL B cells alone were always less than 900 cpm. Data are expressed as the ratio of 3H-TdR incorporation of FL B cells cultured with stromal cells/(3H-TdR incorporation of FL B-cells alone + 3H-TdR incorporation of stromal cells alone).
- Figure S1. Phenotypic characterization of FDCs, Resto, and BM-MSCs (JPG, 72.8 KB)
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(A) FDC-enriched fractions were obtained at the 15%/25% interface of a discontinuous Percoll gradient. Purified tonsil B cells were obtained using the B-cell isolation kit II (Miltenyi Biotech, Bergisch Gladbach, Germany), as the unbound fraction. FDC-enriched fraction, Resto cells, BM-MSCs, and tonsil B cells were fixed in cold acetone and stained with May-Grünwald Giemsa (MGG), FITC-conjugated anti-CD45 mAb (Beckman Coulter, Villepinte, France), or with purified anti-CD21 long isoform (clone DRC-1 DakoCytomation, Glostrup, Denmark), CD21 (Beckman Coulter), and CD90 (Becton Dickinson, San Diego, CA) followed by labeling with specific secondary antibodies conjugated to Fluoprobes 546 (Interchim, Montlucon, France). Arrows indicate CD21+ or CD45+ lymphocytes adherent to FDCs. Original magnification for MGG staining, ×400 for FDCs and B cells, ×100 for Resto cells and BM-MSCs. Bars represent 20 µm. (B) Expression of CD73 (Becton Dickinson) and CD105 (Beckman Coulter) on Resto cells analyzed by flow cytometry. Blue lines indicate immunoglobulin control, red lines indicate specific staining.
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