Chang medium b c

Cardiac tissues were obtained from fetuses at 18–22 weeks of gestation after therapeutic abortion according to protocols approved by our Institutional Ethical Committee. Tissues and RNAs were stored at the Telethon Bank of Fetal Biological Samples at the University of Naples. For this study, 3 hearts from euploid fetuses (NH), 3 from fetuses with DS without CHD (DH), and 3 from fetuses with DS and CHD (CDH) were analyzed. For let-7c mimic and inhibitor transfection, 2 previously characterized [21 (link), 22 (link)] primary lines of euploid (N-HFF) and trisomic (DS-HFF) fetal fibroblasts were, respectively, used. Fibroblasts from biopsies were cultured in T25 flasks (BD Falcon) with Chang Medium B + C (Irvine Scientific) supplemented with 1% penicillin/streptomycin (Gibco) at 37°C in 5% CO₂ atmosphere; all the analyses described throughout this study were carried out at cell culture passages 4-5.

Fibroblasts were cultured in T25 flasks (BD Falcon) with Chang medium B + C (Irvine Scientific) supplemented with 1% penicillin/streptomycin (Gibco) at 37 °C in a 5% CO₂ atmosphere. All analyses described throughout this study were carried out at cell culture passages 4–5.
Analysis of karyotype was performed according to standard methods (Genesio et al., 2011 (link)).