hiPSCs were cultured (Supplementary Figure S1) and in vitro differentiated using an embryoid body (EB) system similar to published protocols.
Peritoneal cells were counted using Turks solution and differentiated using anti-F4/80 (Clone BM8; BioLegend) and anti-Ly6G (Clone 1A8; BioLegend) antibodies.
Furthermore, via weak non-covalent bonding interactions, various peptides may be identified and differentiated using engineered protein pores with different surface functional groups.
To obtain fully differentiated adipocytes, the 3T3-L1 preadipocytes were plated and grown on 10 cm plates to 2 days post confluence and induced to differentiate using a standard induction cocktail of 3-isobutyl-1-methylxanthine, dexamethasone, and insulin (MDI) as previously described [8].
The ES cell line 46C (Sox1-GFP) [ 32] was grown and maintained as previously reported [ 33] and induced to differentiate using a monolayer protocol.
Confluent preadipocytes were induced to differentiate using a standard differentiation medium (DM2-L1, Zen-Bio Inc, Research Triangle Park, NC, USA).
Expanded clones were induced to differentiate using a standard AC protocol with Dmed, and marker expression was evaluated at the level of proteins (SOX9, COL2A1 and COL1A1).
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Two days post-confluence, the cells were induced to differentiate using an adipogenic cocktail (1 µg/ml insulin, 0.5 mM isobutylmethylxanthine, and 1 µM dexamethasone) added to a basal medium (DMEM with 10% FBS and penicillin/streptomycin).
Pre-adipocytes were plated in DMEM with 10% FBS and 1% P/S and then differentiated using the same protocol for 3T3-L1 adipocytes.
