Journal: Breast Cancer Research and Treatment

Article Title: Adipose stem cell crosstalk with chemo-residual breast cancer cells: implications for tumor recurrence

doi: 10.1007/s10549-018-05103-w

Figure Lengend Snippet: Adipose stem cells (ASCs) Migrate toward Conditioned Media (CM) from Chemo-Residual TNBC Cells. A CM was prepared by growing chemo-residual SUM159 TNBC cells in serum-free media for 48 h. The ability of primary human ASCs (Zenbio) to migrate toward this CM (Chemo-residual CM), FBS (+ Control) or serum-free media (− Control) was evaluated in a 15 h transwell assay. Total number of migrated cells from 5 representative fields (100 × magnification) was determined for each well, and mean cell number from triplicate wells (± SD) was calculated. Significance was determined by two tailed t test (**** p < 0.0001). Similar results were obtained in at least three independent trials. B Representative fields (100 ×) showing migration of ASCs towards chemo-residual TNBC CM in a 15 h transwell assay. C CXCR4 expression was assessed by incubating ASCs with CXCR4 antibody (blue line) or control IgG (red line), followed by FITC-conjugated secondary antibody. CXCR4 cell surface expression was determined by flow cytometry. D RNA was isolated from untreated and chemo-residual SUM159 cells and subjected to SDF-1 alpha and GAPDH real-time PCR. SDF-1 alpha gene expression relative to GAPDH is presented for both cell lines

Article Snippet: A CM was prepared by growing chemo-residual SUM159 TNBC cells in serum-free media for 48 h. The ability of primary human ASCs (Zenbio) to migrate toward this CM (Chemo-residual CM), FBS (+ Control) or serum-free media (− Control) was evaluated in a 15 h transwell assay.

Techniques: Control, Transwell Assay, Two Tailed Test, Migration, Expressing, Flow Cytometry, Isolation, Real-time Polymerase Chain Reaction, Gene Expression

Journal: Breast Cancer Research and Treatment

Article Title: Adipose stem cell crosstalk with chemo-residual breast cancer cells: implications for tumor recurrence

doi: 10.1007/s10549-018-05103-w

Figure Lengend Snippet: CXCR4 and SDF-1α Neutralizing Antibodies Block ASC Migration toward chemo-residual TNBC cell Conditioned Media. A Human ASC migration toward chemo-residual TNBC cell CM (prepared as described in Fig. ) was measured in the presence of a CXCR4-neutralizing Ab (EMDMillipore, 10 µg/mL), an SDF1α -neutralizing antibody (R&D Systems, 10 µg/mL), or control IgG (R&D Systems, 10 µg/mL). Chemotaxis was measured as in Fig. . The right panels show representative fields of migrated ASCs in the presence of the indicated antibodies. Significance was measured with a two tailed t test (*** p < 0.0005). B The ability of a CXCR4 small molecule inhibitor (AMD3100; EMD Millipore, 50 µg/mL) to block ASC migration toward chemo-residual TNBC cell CM was tested in a 15 h transwell assay. The bottom panels are representative fields of migrated ASCs ± AMD3100. Significance was measured with a two tailed t test (*** p < 0.0005). C The ability of ASCs to migrate toward chemo-residual TNBC cell CM was tested in a 15 h transwell assay in the presence of the indicated concentrations of CXCR4 small molecule inhibitor (AMD3100). Significance was measured using a two tailed t test (** p < 0.005; *** p < 0.0005)

Article Snippet: A CM was prepared by growing chemo-residual SUM159 TNBC cells in serum-free media for 48 h. The ability of primary human ASCs (Zenbio) to migrate toward this CM (Chemo-residual CM), FBS (+ Control) or serum-free media (− Control) was evaluated in a 15 h transwell assay.

Techniques: Blocking Assay, Migration, Control, Chemotaxis Assay, Two Tailed Test, Transwell Assay

Journal: Breast Cancer Research and Treatment

Article Title: Adipose stem cell crosstalk with chemo-residual breast cancer cells: implications for tumor recurrence

doi: 10.1007/s10549-018-05103-w

Figure Lengend Snippet: ASC CM Increases Proliferation of Chemo-residual TNBC Cells in an FGF2-dependent manner. A and B ASC CM was prepared from human ASCs (ZenBio) grown in reduced serum media for 72 h. This ASC CM (ASC CM), or control reduced serum media (Control) was added to chemo-residual SUM159 ( A ) or chemo-residual BT549 ( B ) cells. The number of chemo-residual TNBC cells was determined after 24 h by trypan blue staining. Results are reported as mean cell number (± SEM) from triplicate wells. Similar results were obtained in 3 trials. Significance was measured using a two tailed t test, *** p < 0.0005. ( C and D ) Chemo-residual SUM159 cells ( C ) or chemo-residual BT549 cells ( D ) were incubated with control media, ASC CM + control IgG, or ASC CM + FGF2-neutralizing antibody (EMD Millipore, 10 µg/mL) for 24 h. Cell numbers were determined as in A. Significance was measured with a two tailed t test (** p < 0.005). This effect was independently observed in three trials. Of note, incubation of chemo-naïve SUM159 cells with ASC CM did not induce cell proliferation (data not shown)

Article Snippet: A CM was prepared by growing chemo-residual SUM159 TNBC cells in serum-free media for 48 h. The ability of primary human ASCs (Zenbio) to migrate toward this CM (Chemo-residual CM), FBS (+ Control) or serum-free media (− Control) was evaluated in a 15 h transwell assay.

Techniques: Control, Staining, Two Tailed Test, Incubation

Journal: Breast Cancer Research and Treatment

Article Title: Adipose stem cell crosstalk with chemo-residual breast cancer cells: implications for tumor recurrence

doi: 10.1007/s10549-018-05103-w

Figure Lengend Snippet: ASC cross-talk with chemo-residual TNBC cells- implications for tumor recurrence. A Chemo-residual TNBC cells secrete the chemokine SDF1α, resulting in recruitment of ASCs, which express the SDF1α chemokine receptor CXCR4, to the tumor site. B ASCs secrete FGF2, which binds to an FGF2 receptor on chemo-residual tumor cells, initiating signaling that drives tumor cell proliferation. Considering that chemo-residual tumor cells can remain dormant in patients for months to years, this ASC/chemo-residual tumor cell cross-talk likely contributes to tumor recurrence in patients post-chemotherapy treatment

Article Snippet: A CM was prepared by growing chemo-residual SUM159 TNBC cells in serum-free media for 48 h. The ability of primary human ASCs (Zenbio) to migrate toward this CM (Chemo-residual CM), FBS (+ Control) or serum-free media (− Control) was evaluated in a 15 h transwell assay.

Techniques: