2.2. Isolation and Identification of Adipo-sEVs

Adipo-sEVs were isolated by ultracentrifugation. Briefly, after HS-5 was fully differentiated into adipocytes, they were rinsed with PBS then cultured in serum-free lipogenetic differentiation medium for 48 h under 37°C. The conditioned medium was collected, and the sEVs were isolated using ultracentrifugation as previously described [12]. Transmission electron microscopy (TEM) was utilized to document the morphology of the sEVs. Nanoparticle analysis (NTA, ZetaView PMX 110) was used to detect the size distribution of the sEVs. The quality of isolated sEVs was analyzed by detecting the expressions of sEVs characteristic markers TSG-101 (Abcam), CD63 (Abcam), and CD9 (Abcam) and non-sEV marker GM130 (Abcam).