American Society of Hematology

Fig. 3.

$Fig. 3. Induction of cell-cycle progression and proliferation of clonal cells after huCD40LT stimulation of primary MCL cell cultures. / Isolated mononuclear cells from 4 MCL patients were seeded at 2 × 106 cells/mL and cultured in the presence or absence of 400 ng/mL for 4 days. On day 4, the cultures were incubated with bromodeoxyuridine (BrdUrd) for 24 hours. After harvest, the cells were analyzed using multiparameter flow cytometry. (A) Dot-plot analyses of kappa and lambda light-chain expression, respectively, combined with detection of BrdUrd incorporation in one MCL cell culture after stimulation with huCD40LT. (B) Dot-plot analyses of simultaneous detection of BrdUrd incorporation (DNA synthesis) and 7-amino-actinomycin-D (7-AAD) (DNA content) after exclusion of small DNA fragments and of cells that did not clearly express the light chain of the malignant clone of the particular MCL case. Dot-plot analyses of 4 huCD40LT-stimulated and -unstimulated MCL cell cultures are shown (displayed dots represent 25% of total counts). The percentages of cells in the framed regions indicate the fraction of cells that traversed through S-phase with BrdUrd incorporation, completed cell division, and returned to G1-phase. sCD40L indicates huCD40LT stimulation.$

Induction of cell-cycle progression and proliferation of clonal cells after huCD40LT stimulation of primary MCL cell cultures.

Isolated mononuclear cells from 4 MCL patients were seeded at 2 × 10⁶ cells/mL and cultured in the presence or absence of 400 ng/mL for 4 days. On day 4, the cultures were incubated with bromodeoxyuridine (BrdUrd) for 24 hours. After harvest, the cells were analyzed using multiparameter flow cytometry. (A) Dot-plot analyses of kappa and lambda light-chain expression, respectively, combined with detection of BrdUrd incorporation in one MCL cell culture after stimulation with huCD40LT. (B) Dot-plot analyses of simultaneous detection of BrdUrd incorporation (DNA synthesis) and 7-amino-actinomycin-D (7-AAD) (DNA content) after exclusion of small DNA fragments and of cells that did not clearly express the light chain of the malignant clone of the particular MCL case. Dot-plot analyses of 4 huCD40LT-stimulated and -unstimulated MCL cell cultures are shown (displayed dots represent 25% of total counts). The percentages of cells in the framed regions indicate the fraction of cells that traversed through S-phase with BrdUrd incorporation, completed cell division, and returned to G₁-phase. sCD40L indicates huCD40LT stimulation.

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