Co-culture Of Adipose Derived Stem Cells And Schwann Cells Leads To Differentiation And Spheroid Formation
Annika Resch, MD, Alexandra-Larissa Stetco, Mac, Tamara Weiss, PhD, Christine Radtke, MD, FEBOPRAS, MBA.
Medical University of Vienna, Vienna, Austria.
Introduction: A spheroid is defined as a self-assembled group of cells representing a 3-D cell cultures. These 3-D cell cultures are known to be more similar to cells in vivo compared to 2-D monolayer cell cultures. Due to enhanced cell to cell interaction and increased secretion of growth factors spheroids show positive effects on differentiation and regenerative processes. Several techniques to generate spheroids have been described in literature so far. The purpose of creating spheroids in vitro is to use and compare their abilities such as enhanced pluripotent potential, increased adipogenesis (PPAR-y), immunomodulatory, proangiogenic (VEGF), antifibrotic and antioxidant cell activity for regenerative and experimental purposes in cell-based research and tissue engineering. Here, human adipose-derived stem-cells (ADSCs) were seeded in co-culture with human Schwann cells to evaluate the effects of Schwann cells on ASCs and their differentiation properties. Methods: Human ADSCs were isolated from the lipoaspirat of healthy patients after undergoing liposuction. Cells were characterized by immunostaining with monoclonal mouse and rabbit antibodies against CD90, CD44, CD34, CD45 as well as stro-1. Immunofluorescence showed positivity for CD90 and CD44 and cells were negative for CD34, CD45 and stro-1. The human Schwann cells where isolated from the ischiadic nerve of an organ donor. In immunocytochemical staining cells were positive for anti-S100 in the immunofluorescence. After isolation and characterization cells were cultured in a T25 flask coated with Laminin/PLL in Schwann cell medium. Results: After about 20 days in co-culture we discovered that cells would detach and proliferate leading to spheroids formation. These findings could only be observed in the co-culture of ADSCs and Schwann cells and were not present in either monoculture of these cells. The 3-D cell aggregates in the supernatant were isolated using the cytospin method. Against all odds immunofluorescence staining showed positivity for S100B suggesting that the spheroids not only consist of ADSCs and fibroblasts but also Schwann cells. Conclusion: Regarding the positive effects of spheroids described in literature so far, our findings might be of significance in our ADSCs and Schwann-cell research on nerve regeneration. Further investigations and control trials are planned to validate our findings including cell characterization via immunofluorescence staining and ELISA for growth factor production.
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