4.2. Isolation of Human Visceral vASCs

Clinical information regarding the donors of vASCs is shown in Table 1. Visceral adipose tissue (VAT) was collected from the intra-abdomen of human donors (n = 5) during gynecologic surgery. The waist-to-hip ratio (WHR) is a strong predictor of visceral obesity [49]. Therefore, based on WHR, vASCs obtained from five donors with visceral obesity (WHR ≥ 0.85) were used. The procedure was approved by the Seoul National University Hospital Institutional Review Board (SNU-1003-009-311). This study was conducted in accordance with the Declaration of Helsinki and informed consent was obtained from the donors for this research. vASCs were isolated as described previously [4]. Briefly, blood vessels of the VAT were removed. VAT was dissociated with collagenase type IA and diluted in 0.25 mg/mL phosphate-buffered saline (PBS) for 1 h at 37 °C, and then centrifuged at 500× g for 4 min. After centrifugation, the stromal vascular fraction (SVF) pellet was collected. The SVF was filtered using MACS through a negative selection of CD31 (endothelial cell marker) and CD45 (hematopoietic stem cell marker). The CD31 and CD45 negative SVF were plated onto a 100 mm culture dish with MesenPRO RS medium containing 1% GlutaMAX and 1% penicillin-streptomycin (PS). After three days of vASCs seeding, non-adherent cells were removed via PBS washing. Expansion of vASCs was performed in a two-passage process before experimental use. vASCs were used for experiments in passages 3–6.

Clinical information regarding the donors of vASCs.

WHR—waist-to-hip ratio; BMI—body mass index.