Cell and culture
This protocol is extracted from research article:
Numerical operations in living cells by programmable RNA devices
Sci Adv, Aug 21, 2019; DOI: 10.1126/sciadv.aax0835

HeLa cells (American Type Culture Collection) were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum (FBS) and 1% antibiotic antimycotic solution (Sigma-Aldrich). Human primary cultured cells were expanded in the provided medium according to the manufacturer’s instructions. NHDFs (Lonza) and NHLFs (Lonza) were cultured with FBM-2 BulletKit containing 2% FBS (Lonza), neonatal NHEKs (Lonza) with KGM-Gold BulletKit (Lonza), and HREs (Lonza) with REGM BulletKit containing 0.5% serum (Lonza). MEFs (Lonza) were expanded in the same medium as HeLa cells. These primary cultured cells were used within 10 passages. hiPSCs derived from NHDFs (201B7) (36) were maintained with StemFit AK03N (AHS) on nippi iMatrix-511–coated well plates. Upon spontaneous differentiation (26), 3000 hiPSCs were seeded on iMatrix-511–coated six-well plates and cultured in StemFit AK03N without bFGF for 14 days. Cell culture without bFGF for 1, 3, and 6 or 9 days was started with 50,000, 10,000, and 3000 hiPSCs, respectively, on 24-well plates. The medium for hiPSCs was changed every second day.

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