Journal: PLoS ONE

Article Title: Glycogen Synthase Kinase 3 (GSK3) Inhibitor, SB-216763, Promotes Pluripotency in Mouse Embryonic Stem Cells

doi: 10.1371/journal.pone.0039329

Figure Lengend Snippet: J1 mESCs co-cultured with MEFs were maintained continuously with vehicle (0.1% DMSO), 1,000 U/ml LIF, 10 or 20 µM SB-216763, 2 µM BIO, or 3 µM CHIR-99021. The cultures were passaged in parallel and 20,000–50,000 mESCs were plated on new MEF-covered 6-well plates every 3 days. A duplicate plate was stained for alkaline phosphatase (AP) activity at days 10, 20, and 30. Shown are representative images from one of two independent trials. Day 60 images were obtained from an independent culture maintained with vehicle (0.1% DMSO), 1,000 U/mL LIF, or 10 µM SB-216763. AP staining on day 60 was carried out on samples from a single trial. The scale bar shown in the day 10 vehicle image represents 100 µm and all images are presented at the same scale.

Article Snippet: CF-1 mouse embryonic fibroblasts (MEF; SCRC-1040, ATCC) were used as a feeder layer for mESC cell culture.

Techniques: Cell Culture, Staining, Activity Assay