Can Extent Of Adipose Tissue Washing Impact The Therapeutic Efficacy Of Autologous SVF Cell Therapies?
Adam J. Katz, MD, Gabriela Aguilo-Seara, BS, Hulan Shang, MS, Scott Northrup, BS, Ramon Llull, MD, PhD.
Wake Forest University, Winston Salem, NC, USA.
PURPOSE: Adipose-derived SVF cell therapies have entered Phase 3 FDA testing. A major challenge for point-of-care autologous cell strategies is the inherent variability that exists when using primary cells derived from a variety of patients. Our central hypothesis is that 1) the extent of adipose tissue washing influences the number of blood derived cells (BDCs) in SVF preparations, and 2) BDCs within SVF preparations confer modifiable effects on SVF bioactivity.
METHODS: Lipo-harvested adipose tissue was divided into equal volumes. One sample was 'washed' by gravity decantation, one by agitation and sump drain irrigation. SVF cells were then isolated using identical methods and the resulting types and numbers of BDCs were characterized by flow cytometry (n=5) . Separately, freshly isolated SVF cells and second passage ASCs were cultured in media containing RBCs or lysed RBCs for up to 7 days (n=4). Cell proliferation was evaluated and conditioned medium was collected on days 0, 3, and, 7 and evaluated for RBC toxins using ELISA.
RESULTS: Tissue washing reduced RBCs nearly fourfold (p=0.004) and leukocytes nearly twofold (p=0.001) compared to decanted samples, while yielding higher Type 2 macrophages (p=0.01). SVF and ASC cell proliferation was higher (2.5-4x) in media with intact RBCs (p=0.001) and was inversely related to levels of free hemoglobin and hemin.
CONCLUSION: The extent of washing of adipose tissue prior to cell isolation can significantly impact the final cell composition of SVF therapies. These differences in composition can impact biological activity and may influence therapeutic efficacy.
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