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Additional Supporting Information may be found in the online version of this article.

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STEM_787_sm_supplfigure1.pdf484KSupplemental Figure 1. Characterization of human and murine MSCs. (A) hMSCs at passage 2 showed typical fibroblast like morphology, and were positive for CD90, CD166, CD73 and negative for CD45. When plated in corresponding differentiation media, hMSCs were capable of differentiating into adipocytes, osteoblasts and chondrocytes, evaluated respectively by Oil Red O staining (×200), Alizarin Red S staining (×100) and collagen type II staining (×100). (B) Using our optimized protocol, mMSCs already appeared morphologically homogeneous at passage 2. They were positive for CD105, CD90, Sca-1, lacked expression of CD45, and also exhibited tri-lineages differentiation potentials. Representative pictures of five independent experiments are shown. Scale bar=100μm. Plots show isotype control IgG staining profile (red) versus specific Ab staining profile (white)
STEM_787_sm_supplfigure2.pdf131KSupplemental Figure 2. Migration of mMSCs towards lung and liver over time. After inoculation of mMSCs i.v. in naive and 5T33MM mice, the cell number in the lungs gradually decreased, while it increased in the liver.
STEM_787_sm_supplfigure3.tif2110KSupplemental Figure 3. Murine 5T33MMvv and 5T33MMvt cells were both positive for CCL25 by PCR analysis.
STEM_787_sm_supplfigure4.pdf300KSupplemental Figure 4. MSCs favor the proliferation of MM cells in vivo. 5T33MMvv cells (5×106) were injected intrafemorally with or without mMSCs (5×105). One week later, more MM cells were in a proliferative state as shown by positive PCNA expression in the femur where mMSCs were co-injected compared to tumor cell injection alone. The slides were counterstained by DAPI. One representative photo is shown. The scale bar=100μm.
STEM_787_sm_supplfigure5.pdf282KSupplemental Figure 5. Minimally detectable MSCs were present in the MM cell population after separation from cell-cell contact coculture. (A) More than 99% hMSCs are positive for CD90. (B) RPMI8226 cells are negative for CD90 (middle). After 48h of cell-cell contact coculture (MSCs: MM=1:10), MM cells were collected by gently pipetting to separate them from adherent hMSCs, and used to test apoptosis and indicated protein expression. In the collected MM cell fraction, there was minimal contamination of detectable MSCs (right).
STEM_787_sm_supplfigure6.pdf624KSupplemental Figure 6. MSCs protect MM cells against apoptosis in vivo. A. MSCs protected MM cells against spontaneous apoptosis in vivo. 5T33MMvv cells (5×106) were injected intrafemorally with or without mMSCs (5×105). One week later, there were less MM cells undergoing apoptosis in the femur where mMSCs were co-injected, compared to tumor cell injection alone, although this was not significant. TUNEL immunofluorescence staining was performed, and the slides were counterstained by DAPI. One representative photo is shown. The scale bar=100μm. B. MSCs protected MM cells against Bortezomib induced apoptosis in vivo. After 5T33MMvv injection into naive mice intrafemorally, five doses of Bortezomib at 0.6mg/kg i.p. every other day were given, leading to MM cell apoptosis as measured by TUNEL staining. However, with the co-injection of mMSCs, significantly less MM cells underwent apoptosis. One representative photo is shown. The scale bar=100μm.

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