Il 23

We performed evaluations on soluble factors from supernatants of uncultured mSVF and eSVF through the commercially enzyme-linked immunosorbent tests (ELISA). To this end, mSVF and eSVF were centrifuged at 600 g for 10 min, and their supernatants were collected to test the released cytokines. Moreover, we plated and cultured mSVF and eSVF to test whether the in vitro culture can modify their cytokine profile compared to the native uncultured adipose products. After 80% confluence, culture supernatants (S/N) from both mSVF (mSVF S/N) and eSVF (eSVF S/N) were recovered and kept in serum-free medium for 48 h before their collection. We performed ELISA assessments on the following molecules: vascular endothelial growth factor (VEGF) (Cloud-Clone Corp, Texas, US), Platelet-derived growth factor (PDGF)-bb (LifeSpan Biosciences, Washington, US), insulin growth factor (IGF)-1 (Cloud-Clone Corp), hepatocyte growth factor (HGF) (Cloud-Clone Corp), interleukin (IL)-10 (Cloud-Clone Corp), interleukin (IL)-1β (Cloud-Clone Corp), IL-6 (Cloud-Clone Corp), IL-21 (MyBioSource, California, US), IL-23 (MyBioSource), tumour necrosis factor (TNF)-α (Cusabio, Houston, Texas, US). We used the manufacturer’s instructions through spectrophotometer measurement at a wavelength of 510–570 nm.

Brain natriuretic peptide (BNP) as a diagnostic and prognostic marker of HF was measured in serum using ELISA kit (MyBioSource, Inc. San Diego, CA, United States). In addition, the following parameters were estimated in tissue homogenates using the corresponding ELISA kits; IL-17 (Sinogeneclon Co., Ltd., China), IL-6 (Chongqing Biospes Co., Ltd., China), P-STAT-3 (Creative Diagnostics; NY, United States), Bax, Bcl2, IL-23, TGF-β, PAF, and atrogin-1 (MyBioSource, Inc. San Diego, CA, United States). A specific ELISA Kit (ab176657, Cambridge, MA, United States) were used to measure both p-Akt (Ser473) and total Akt in tissue homogenate.