Journal: Scientific Reports

Article Title: Improving the differentiation potential of pluripotent stem cells by optimizing culture conditions

doi: 10.1038/s41598-022-18400-8

Figure Lengend Snippet: The differentiation potential of cells in culture can be altered by culture medium. ( A ) H9 cells cultured with Essential 8 (Es8) medium on vitronectin-N (VTN)–coated dishes were transferred to RFF2 medium, cultured for 15 days (3 days/passage × 5 passages), transferred again to Es8 medium, cultured 24 days (3 days/passage × 8 passages), and then transferred again to RFF2 medium. Photos of cells in designated culture conditions, with the cell number scored at day 3 after seeding 1.0 × 10 5 cells (left panels); flow cytometric analysis of CHD7, CHD7 copy numbers from 5 ng total RNA at day 3 (middle panels); and photographs of EBs formed by day 14 from cells in each culture condition and numbers of EBs formed (right panels). The results are representative of three independent experiments. ( B ) H9 cells were cultured either with Es8 or RFF2 on VTN-N–coated dishes. The loci of copy number variants (CNVs) detected when cells were cultured with Es8 medium (left panels) or RFF2 medium (right panels) are shown. CHD7 expression was determined by flow cytometry (mean values are shown), and CHD7 copy numbers were determined by digital droplet PCR in cells cultured with Es8 or RFF2 medium.

Article Snippet: Cells in single-cell suspensions were seeded at 1 × 10 5 cells/well in 6-well plates when cultured with Essential 8 (Es8) medium and 1–3 × 10 5 cells/well when cultured with Repro FF2 (RFF2) medium (ReproCELL; 4 mL/well).

Techniques: Cell Culture, Expressing, Flow Cytometry