Plastic Surgery Research Council
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PSRC 60th Annual Meeting
Program and Abstracts

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Adipose-derived Stem Cells after Paclitaxel Treatment demonstrate decreased Function and Suppression of Breast Cancer Cell Viability
William M. Harris, M.D., Michael A. Plastini, MD, Telisha Ortiz, MD, Nikolas Kappy, MD, Jefferson Benites, BS, Shaohua Chang, PhD, A. Lelani Fahey, MD, Martha S. Matthews, MD, Jeffrey P. Carpeneter, MD, Spencer Brown, PhD, Ping Zhang, PhD.
Department of Surgery- Cooper University Hospital and Cooper Medical School of Rowan University, Camden, NJ, USA.

PURPOSE: Stromal vascular fraction (SVF) is used in breast reconstruction in FDA clinically approved studies. In SVF, adipose derived stem cells (ASC) may play a role in wound healing. In patients following chemotherapy for breast cancer, poor soft tissue wound healing is a major problem. Previous work in our laboratory demonstrated that direct exposure to Paclitaxel (PTX) has cytotoxic effects on ASCs by decreasing proliferation and multipotency. This study examined human ASCs pretreated with PTX and then exposed to a washout period (no PTX) of 8 days to evaluate 1) the influence on ASC function after chemotherapy cessation and 2) influence on tumor cell growth in ASC co-culture.
METHODS: IRB-approved human ASCs were isolated and MDA-MB-231 cells (breast cancer cell line) were treated with PTX at different concentrations. Cell proliferation, viability, and migration were measured by growth curves, MTT assays, and scratch assays. ASCs were cultured in derivative-specific differentiation media with or without PTX (1uM) for 3 weeks. Adipogenic and endothelial differentiation were measured by quantitative RT-PCR, Oil-Red-O staining and cord formation on Matrigel, respectively. MDA-MB-231 cells and ASCs were co-cultured at equal cell numbers. ASC conditional media were collected every 3 days from ASCs, both no-PTX treatment control and PTX treated - washout groups.
RESULTS: Following cessation and washout of PTX, 2.4-fold (0.1uM) and 4.0-fold (1uM) lower proliferation rates were observed in ASC when compared with a no-PTX treatment control group. In comparison with no-PTX untreated controls, PTX treated - washout ASC had decreased cell migration (relative scratch width: 29±10% vs. 86±4%, p<0.05). PTX treated - washout ASCs compared to no-PTX ASC treatment group demonstrated: 1) decreased rates of adipogenic differentiation with lower Oil-Red-O positive stained lipid droplets and decreased mRNA levels of PPAR-γ, LPL; and 2) decreased rates of endothelial differentiation with decreased cord formation on Matrigel with decreased levels of CD31, vWF and eNOS. In a second series of studies, MDA-MB-231 cells treated with PTX treated - washout ASC conditional medium (ASC-CM) for 8 days (indirect co-culture) exhibited a decrease in cell growth (10 fold in PTX pre-treated CM; p<0.01). The cumulative population doubling time was longer for the ASC-CM (PTX pre-treated) compared to conditional medium from ASC with no exposure to PTX (5.89±0.38 day vs. 1.61±0.04 day, p<0.01). In direct co-culture with both ASC and MDA-MB-231, total cellular mRNA expression of Caspase 3 (1.7 fold, p<0.01) was increased while mRNA expression of the breast cancer marker SNCG (1.6 fold, P<0.05) was decreased compared to respective cell types cultured separately for 7 days.
CONCLUSION: Our results showed that ASCs after PTX treated - washout had decreased proliferation and multi-lineage differentiation rates. Direct ASC contact and conditional medium from PTX treated - washout ASCs suppressed tumor cell growth. These results may provide insight into associated poor wound healing in cancer patients following Paclitaxel cessation. If ASCs function is altered by PTX exposure and is associated with poor post-surgical wound healing, then the isolation, banking and use of autologous adipose SVF may be considered to minimize wound adverse events.


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