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. 2011 Apr;25(4):707-11.
doi: 10.1038/leu.2010.300. Epub 2011 Jan 14.

A unique three-dimensional SCID-polymeric scaffold (SCID-synth-hu) model for in vivo expansion of human primary multiple myeloma cells

T Calimeri, E Battista, F Conforti, P Neri, M T Di Martino, M Rossi, U Foresta, E Piro, F Ferrara, A Amorosi, N Bahlis, K C Anderson, N Munshi, P Tagliaferri, F Causa, P Tassone
Free PMC article

A unique three-dimensional SCID-polymeric scaffold (SCID-synth-hu) model for in vivo expansion of human primary multiple myeloma cells

T Calimeri et al. Leukemia. 2011 Apr.
Free PMC article
No abstract available

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Figures

Figure 1
Figure 1
(A) SEM analysis of a PCLS ( × 65 (1), × 500 (2), × 650 (3) and × 1200 (4) magnification) shows interconnected large and small pores (white bars=1 mm (1), 200 μm (2), 100 μm (3) and 50 μm (4)). (B) Comparative SEM analysis ( × 150 magnification) between a synthetic PCLS (upper panel) and a surgical sample of human femur adult bone (lower panel) shows similar microarchitecture (white bars=500 μm). Surface morphology was studied by a Leica Cambridge (Stereoscan S440) SEM (Cambridge, UK) at an accelerating potential of 20 kV. (C) H&E staining of adherent OP9 mouse stromal cells at 3 weeks after scaffold implant in mice. (D–F) SEM analysis ( × 3500 magnification) (d), H&E staining (e) and confocal laser scanning microscopy (f) show adherence within a PCLS of human BMSCs, obtained from two different patients (pt #3, (a); pt #7, (b) See Table 1), 1 week after implant in mice. (G) Flow chart diagram. Schematic representation of experimental methodology.
Figure 2
Figure 2
(a) Histological and immunohistochemical (anti-CD138/MI15 (Dako, Dako Itala S.p.A., Milano, Italy), anti-κ (Dako) and anti-λ (Dako)) analysis of retrieved PCLSs. Line 1 shows unselected BMMCs; line 2 shows selected CD138+ on heterologous BMSCs; line 3 shows selected CD138+ from PB of a PCL patient on heterologous BMSCs; and line 4 shows IL-6-dependent INA-6 MM cells on heterologous BMSCs. Anti-human κ-light chains and anti-human λ-light chains were used (Dako). Dilutions were 1:15 000 for κ-light chains and 1:10 000 for λ-light chains. (b) Ki-67 staining of MM-bearing PCLSs retrieved 3–4 months after cell injection. (c, d) Kinetics of appearance of paraproteins in sera from representative SCID-synth-hu mice engrafted with samples from MM patients. A code for each patient sample is provided according to Table 1. Left panel includes seven samples from patients with whole paraprotein (ELISA, GenWay Biotech Inc., San Diego, CA, USA); right panel includes two patients with light chain MM (ELISA, Bethyl Laboratories Inc., Montgomery, TX, USA).
Figure 3
Figure 3
(a) SCID-synth-hu mice were treated intraperitoneally with bortezomib (1 mg/kg)+dexamethasone (1 mg/kg). The black arrows indicate the time of treatment. (b) H&E staining of a MM-bearing PCLS, engrafted with primary MM cells, following Bort+Dex treatment showed diffuse stromal and intracellular calcification demonstrating massive apoptosis. (c) (1) H&E staining of a retrieved PCLS showed vessels of variable size (black circles). (2) Immunohistochemistry of a PCLS coated with BMSCs demonstrated reactivity with an anti-CD31/JC70A (Dako) specific for human CD31 (black arrow). Appropriate positive control tissues were added on each automated immunohistochemistry run to confirm antibody specificity. Negative control section included normal rabbit serum as a substitute for the primary antibody. (3) H&E staining of a retrieved PCLS engrafted with BMMCs shows an active site of vasculogenesis within an area of MM cell growth (yellow arrow). (4) H&E staining of a retrieved PCLS coated with BMSCs revealed the presence of a neosynthesized ECM inside the pores.
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