Figure 2.
(A) Jeko, SP53 and Z138 MCL cells (5×103) were cultured in PHA-LCM (phytohemagglutinin human leukocyte conditioned medium) methylcellulose medium. To test the role of the CXCR4/SDF-1 signaling axis in colony formation in PHA-LCM medium, SDF-1 (200 ng/ml) or AMD3100 (40 μM) or AMD3100 with SDF-1 were added during culture. Cells were counted as a colony if they had more than 40 cells. Colony numbers were combined from triplicate plates.
(B) MCL cells were cultured in PHA-LCM medium with human bone marrow stromal cells, HS27a, or SDF-1 neutralizing antibodies (100 mg/ml). In some experiments, MCL cells were also cultured using stromal cells for 24 hours and collected for the colony forming assays.