Journal of Oncology / 2012 / Article / Fig 4

Research Article

Chronic Myelogenous Leukemia Cells Contribute to the Stromal Myofibroblasts in Leukemic NOD/SCID Mouse In Vivo

Figure 4

Functional analysis of the separated BCR-ABL positive myofibroblasts from CML cell-engrafted NOD/SCID murine bone marrow. (a) RT-PCR analysis of the indicated cells on VEGF-A system. (b) ELISA assay of human VEGF-A in the culturing supernatants. The values represent the mean % ( 𝑛 = 3 ) Β± SD. SCID 𝑛 -Fib, myofibroblasts from NOD/SCID murine bone marrow; Hu 𝑛 -Fib, normal human bone-marrow-derived myofibroblasts; Whole-Fib, CML cell-derived myofibroblasts including BCR-ABL positive and negative cells that were prepared in vitro; (βˆ’)-Fib, CML bone marrow-derived BCR-ABL negative myofibroblasts prepared in vitro; (+)-Fib, CML bone marrow-derived BCR-ABL positive myofibroblasts prepared in vitro; SCID (+)-Fib, CML cell-engrafted NOD/SCID murine bone-marrow-derived BCR-ABL positive myofibroblasts. The asterisk of (+)-Fib indicates 𝑃 < 0 . 0 1 between (+)-Fibs and other left side 4 Fibs, and the asterisk of SCID (+)-Fib indicates 𝑃 < 0 . 0 1 between SCID (+)-Fibs and other left side 5 Fibs. (c) Cellular morphology when co-cultured with myofibroblasts. 1: Parental CML cells were cultured on the control NOD/SCID murine bone- bone marrow-derived myofibroblasts. 2: The parental CML AP cells were cultured on CML cell-engrafted NOD/SCID murine bone marrow-derived BCR-ABL-carrying SCID (+)-Fibs. 3: Normal bone marrow non-adherent CD34-positive cells were cultured on SCID (+)-Fibs (Γ—200). (d) Cell-proliferation assay. 1–3: normal bone marrow-derived non-adherent CD34-positive cells, and 4–8: parental CML AP cells. 1, 4: No feeder layers, 2, 5: NOD/SCID murine bone marrow-derived stromal myofibroblasts, and 3, 6–8: CML cell-engrafted NOD/SCID murine bone marrow-derived BCR-ABL positive myofibroblasts. 7: Cultures are added with anti-human VEGF-A Ab, and 8: with control Ab. Asterisks indicate 𝑃 < 0 . 0 1 .

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