Anti cd105 antibody

After expansion of each group of riPSCs, they were compacted to form EBs in hanging drops for 2 days. Over the following 3 days, they were exposed to 1×10^-6 M RA in culture medium followed by washing for 1 day without RA. After 6 days, the EBs were plated onto gelatin-coated dishes and then incubated with 850 nM insulin (Sigma) and 20 nM 3,3,5-triiodo-l-thyronine (T3; Sigma) in basic-DMEM. MSCs expressing CD105 in developing EBs were sorted by MACS by using both an anti-CD105 antibody (R&D Systems) and an anti-rat IgG antibody conjugated with magnetic beads (Miltenyi Biotec). The sample preparation, magnetic labeling, and magnetic separation with mini-MS columns were conducted according to the manufacturer's instructions. After MACS separation, CD105+ MSCs were transferred to gelatin-coated dishes. For further induction to adipocytes, they were maintained in 850 nM insulin, 20 nM T3, and 1 µM dexamethasone (Dex; D8779, Sigma) in basic DMEM. In addition, 16 µM recombinant rat leptin (400-21; PeproTech, Rocky Hill, NJ) was supplemented for some MSCs from both groups of rat iPSCs, i.e., o-riPSCs and l-riPSCs. The culture medium was changed every day.