Figure 1.
Morphology of primary malignant cells and VR09 cell line.
Morphological features of patient’s primary cells and VR09 cells in suspension, as assessed by May-Grünwald-Giemsa staining. (A) Patient’s peripheral blood cells. (B) Patient’s bone marrow cells. (C) VR09 cells: small and round clumps in suspension. (D) VR09 cells: plasmacytoid appearance, with irregular nucleus, compact chromatin and abundant basophilic cytoplasm.
Table 1.
Primers used to amplify and sequence P53, Card 11, CD79B, MYD88 AND RPMS1 genes.
Table 2.
Marker expression by VR09 cell line and tumor masses, as assessed by immunohistochemistry performed on FFPE cell block.
Figure 2.
Immunophenotyping of VR09 cell line (grey and filled curves) and cell suspensions from in vivo tumor mass (black and filled curves), as compared with isotype control (white curves).
Figure 3.
A) Spherical subcutaneous mass (arrow) in Rag-2−/− γ-chain−/− mice 36 days after VR09 cell line injection (representative case). (B) Timing of tumor development in vivo (6 mice s.c. injected with 5×106 cells/mouse).
Figure 4.
Representative histological markers detected on tumors.
High magnification (20X) of histological sections of tumors developing after in vivo injection of VR09 cell line. Lymphoid infiltrates display large size, plasmablastic-plasmacytic features and high Ki-67 index. Cells express CD19, CD20, CD138, CD79a, IgM, IgG and EBV protein (EBER).
Figure 5.
EBV positivity in the original tumor and in the VR09 cell line.
PCR products analysis by Agilent 2100 Bioanalyzer showed the presence of the same 151 bp specific amplicon for EBV RPMS1 gene, thus demonstrating that EBV infection was present in the original cells from patient. A normal DNA from pancreas was used as negative control.
Figure 6.
Karyotyping and FISH of secondary culture.
(A) Trisomy of chromosome 12 and structural chromosome aberration involving chromosomes 7 and 9 on secondary culture detected by karyotype; (B) Chromosome 9 staining by FISH of secondary culture.
Table 3.
Summary of VR09 cell line features.