Insulin growth factor 1 igf 1

Fibroblast cells were reprogrammed into iPSCs, as previously described [18 (link)]. PPCs were obtained after following a 2D differentiation protocol [37 (link)]. Briefly, iPSCs were dissociated with ReLeSR (Stemcell Technologies) and plated in 12-well plates coated with matrigel (Corning, Tewksbury, MA, USA) to form a monolayer. Essential-Flex E8 medium was changed to differentiation medium (CI) when reaching confluence. The CI medium consisted of DMEM/F12 supplemented with nonessential amino acids (NEAA, Sigma Aldrich, Saint Louis, CA, USA), B27 supplements (Thermo Fisher Scientific, Waltham, MA, USA), N2 supplements (Thermo Fisher Scientific), 100 ng/µL of insulin growth factor-1 (IGF-1, Sigma Aldrich), 10 ng/µL of recombinant fibroblast growth factor basic (bFGF, Sigma Aldrich), 10 µg/µL of Heparin (Sigma Aldrich), 200 µg/mL of recombinant human COCO (R&D Systems, Minneapolis, MN, USA), and 100 µg/mL of Primocin (Invivogen, Toulouse, France). Half of the medium was replaced every day for 30 days. On day 28, PPCs were treated with 1 µM of AON. AONs were first mixed with the medium without any transfection reagent and were subsequently added to the cells. Twenty-four hours later, cycloheximide (CHX) was added to the medium (final concentration 100 µg/mL), and on day 30 (48 h post-AON delivery and 24 h post-CHX treatment), cells were collected.