Isolation of Rat Adipose-Derived Stromal Vascular Fraction

SVF cells were isolated from healthy female Sprague-Dawley rats as described elsewhere [9 (link)]. In summary, fat was minced in 0.2% collagenase type I (Thermo Fisher Scientific; #17100017) in HBSS and placed in a 37 °C water bath for 60 min. The enzyme solution was neutralized using Minimum Essential Medium (MEM; Thermo Fisher Scientific; #32561029) supplemented with 10% FBS and filtered through a 70-μm cell strainer. Then, red blood cells were lysed using Ammonium-Chloride-Potassium (ACK) lysing buffer (Thermo Fisher Scientific; #A1049201) for 3 min. ACK buffer was removed by washing/centrifugation and the resulting SVF pellet was resuspended in MEM supplemented with 10% FBS and 1% penicillin-streptomycin.