Joris van Dongen

131 Isolation of SVF by the FAT procedure c. 49.5 ml of demiwater. d. 0.5 ml of acetic acid (glacial). 6. Phosphomolybdic-phosphotungstic acid: a. 2.5 g of phosphomolybdic acid. b. 2.5 g of phosphotungstic acid. c. 50 ml of demiwwater. 7. Aniline Blue: a. 1.3 g of aniline blue. b. 1 ml of acetic acid (glacial). c. 49 ml of demiwater. 8. 1% Acetic acid: a. 0.5 ml of acetic acid (glacial). b. 49.5 ml of demiwater. OPTIONAL: Enzymatic solation of cellular SVF (cSVF) derived from tSVF 1. 0.1% bacterial collagenase A solution: a. 50 mg bacterial collagenase A. b. 50 ml PBS/1% BSA. 2. 100 µm filters (Greiner Bio-one International GmbH, Kremsmünster, Austria). 3. Lymphoprep (Lucron products). 4. Lysis buffer (Pharmacy University Medical Center Groningen, Groningen, the Netherlands). 5. FACS buffer (PBS/0.5% BSA) METHODS Liposuction of adipose tissue 1. Make a small stab incision of 1 cm on the donor site (preferably legs or abdomen). 2. Infiltrate the donor site with a modified Klein’s solution (per 500 ml of saline, 20 ml of lidocaine, 2% epinephrine 1: 200,000 and 2 ml of bicarbonate).