Ix71 imaging system

sf9 cells were transfected with different plasmid as described in the previous section and were imaged at 48 h post-transfection with an Olympus IX71 imaging system using an RFP light cube or bright field.

The mesenchymal stem cell C3H10T1/2 was purchased from Procell Life Science & Technology Co. Ltd. (Wuhan, China). This cell shows fibroblast morphology and is functionally like mesenchymal stem cells (Pinney and Emerson, 1989 (link)). The C3H10T1/2 cells were maintained in minimum essential medium (MEM, HyClone, China) with 10% fetal bovine serum (FBS, Cell-max, Australia; F8318) in a 5% CO₂ environment at 37°C. The medium containing 10% FBS, 125 nM indomethacin, 0.5 mM 3-isobutyl-1-methylxanthine, 1 μM rosiglitazone, 850 nM insulin, 1 nM triiodothyronine, and 1 μM dexamethasone was used for induction of adipocyte directional differentiation. Preadipocytes were cultured in the induced medium for 48 h to induce directional differentiation. These cells were then shifted to the medium with 1 μM rosiglitazone, 1 nM triiodothyronine, 850 nM insulin, and 10% FBS after 48 h and were harvested until the sixth day (Patrick et al., 2011 (link); Zhang et al., 2013 (link)). Subsequently, as observed using an Olympus IX71 imaging system (Olympus, Japan), at 200 X magnifications, the cells became round and exhibited lipid droplets. After cell differentiation, accumulated fats were stained using oil red (Dingguo Changsheng, China; A12989), and optical density was measured at 490 nm with a microplate reader.