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Establishment and characterization of a novel Waldenström macroglobulinemia cell line, MWCL-1

Lucy S. Hodge, Anne J. Novak, Deanna M. Grote, Esteban Braggio, Rhett P. Ketterling, Michelle K. Manske, Tammy L. Price Troska, Steven C. Ziesmer, Rafael Fonseca, Thomas E. Witzig, William G. Morice, Morie A. Gertz and Stephen M. Ansell

Data supplements

Article Figures & Data

Figures

  • Figure 1

    Sequence alignment demonstrating homology between IgVH3–15 and the heavy chain genes of MWCL-1 cells and the primary tumor. PCR-amplified DNA was prepared and sequenced as discussed in “Sequence analysis of IGHV.” The sequence was compared with germline configurations using IMGT/V-QUEST software Version 3.2.17 on the IMGT website (www.imgt.cines.fr/IMGT_vquest/share/textes). Identical VH sequences were obtained for both MWCL-1 cells and the primary tumor, but this common sequence is only referred to as MWCL-1 in the figure.

  • Figure 2

    MWCL-1 cell line morphology. (A) Hematoxylin and eosin staining demonstrating the heterogeneity of the WM tumor clone represented in MWCL-1 cells, including B lymphocytes, lymphoplasmacytic cells, and mature plasma cells (original magnification ×200, Olympic Provus AX70 microscope). (B) Immunofluorescence showing the presence of κ-light chain on the extracellular surface of MWCL-1 cells and cytoplasmic IgM.

  • Figure 3

    Immunophenotypic characterization of the MWCL-1 cell line. FITC-, allophycocyanin-, and phycoerythrin-conjugated antibodies were used for phenotyping of the MWCL-1 cell line by flow cytometry. Blue peaks represents isotype staining; and red peaks, antigen-specific staining.

  • Figure 4

    Whole-genome aCGH profiles. Comparative chromosomal CNAs in the original WM patient sample and the established MWCL-1 cell line at a 6-month passage. Note the common CNAs on chromosome 17 between all samples. Chromosomes 1 through 22, X, and Y, from p to q arm, are plotted on the x-axis from left to right. The predicted number of copies of each gene is indicated to the left of each panel.

  • Figure 5

    Primary WM tumor and MWCL-1 cell line share common chromosome CNAs. Whole chromosome plot of chromosome 17 is shown for the original WM patient sample and the established MWCL-1 cell line at 6 months, illustrating the interstitial losses in 17p13-p13.2 and 17q25.1-q25.3. The predicted number of copies of each gene is indicated on the top of each panel.

  • Figure 6

    MWCL-1 cells retain both copies of chromosome 6q but exhibit monoallelic expression of mutated TP53. (A) Representative FISH image of MWCL-1 cells at a 12-month passage confirming the absence of a 6q deletion. Two hybridization signals are observed for both the probe specific to the centromere of chromosome 6 (green signal), and the locus-specific, c-MYB probe (red signal). (B) Representative FISH image of MWCL-1 cells at a 12-month passage demonstrating monoallelic loss of TP53. Two hybridization signals are observed for the probe specific to chromosome 17 (green signal), but only one signal is observed for the locus-specific, TP53 probe (red signal).

  • Figure 7

    IgM secretion and proliferation of MWCL-1 cells in response to cytokines. MWCL-1 cells were cultured with the indicated cytokines for 72 hours, at which time IgM secretion was quantified by enzyme-linked immunosorbent assay (A), and DNA synthesis measured by 3H-TdR incorporation (B). Bars represent the mean of triplicate experiments ± SD. *Value significantly higher than in the respective untreated control cells (P < .05).