Isolation and Characterization of hMSCs

Human MSCs (hMSCs) were isolated from adipose tissue biopsies, and then cultured, expanded, characterized, and cryopreserved by staff at the Mayo Clinic Human Cellular Therapy Laboratory as previously described in [87 (link),88 (link)]. The release criteria included culture sterility, mycoplasma testing, and cytogenetic analysis, as well as cellular phenotyping, including CD90+, CD105+. CD73+, HLA Class I+, CD14+, CD44+, HLA DR, and CD45-. Cells were cultured in advance MEM media (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with human platelet lysate (5%; Lawson, Mill Creek, WA, USA), 100 U/mL Antibiotic-Antimycotic (Gibco Thermo Fisher Scientific, USA), 100 units/mL Glutamax (Gibco Thermo Fisher Scientific, USA), and 1000 USP units heparin (Novaplus, Lake Zurich, IL, USA) at 5% CO₂ and 37 °C.