Figure 4.

Functional analysis of ex vivo-differentiated primary adipocytes treated with clozapine. SVF-derived hADMSCs were differentiated for 2 weeks to white (W) or positive control beige (Be) adipocytes. Clozapine (red bars) was administered at 100 ng ml⁻¹ concentration on the last 2 and 4 days or during the whole adipogenic differentiation process. (a) Relative mitochondrial DNA amount of human adipocytes determined by quantitative PCR in five different donors. (b) Oxygen consumption of SVF-derived adipocytes of one representative donor measured with an XF96 oxymeter. After recording the baseline oxygen consumption, the cells received a single bolus dose of dibutyril-cAMP. Then, stimulated oxygen consumption was recorded at every 30 min. Proton leak respiration was determined after adding oligomycin to block ATP synthase activity. (c) Basal, cAMP-stimulated and oligomycin-inhibited oxygen consumption levels (as compared with the basal OCR of white adipocytes) in four different SVF-derived adipocyte donors. (d) Effect of short-term cAMP treatment on the expression of UCP1 gene in primary adipocytes. The experiment was repeated five times with SVFs from independent healthy donors (relative gene expression was determined by RT-qPCR, target gene was normalized to GAPDH); *P<0.05, **P<0.01. hADMSC, human adipose-derived mesenchymal stem cell; OCR, oxygen consumption rate; RT-qPCR, quantitative PCR with reverse transcription; SVF, stromal-vascular fraction.