Hyclone rpmi 1640 medium

Cas9 was expressed in HL60 cells, kindly provided by Associate Prof. Mattias Magnusson, through transduction of a modified version of the pLCv2 vector containing a Cas9-P2A-BFP cassette. Transduced cells were then selected based on fluorescent marker expression. The Cas9-expressing HL60 cells were cultured in HyClone RPMI-1640 Medium (Cytiva) supplemented with 10% heat-inactivated FBS (Cytiva) and 1% Penicillin–Streptomycin (Cytiva). Primary CD34⁺ HSPCs were thawed and cultured in Serum-Free Expansion Medium (SFEM, Stem Cell Technologies) complemented with 1% Penicillin–Streptomycin (Cytiva) and Stem cell factor (SCF), FLT3-ligand (FLT3L) and thrombopoietin (TPO) were added in the concentration 100 ng/ml and acquired from PeproTech or Miltenyi Biotec. The CD34⁺ cells were cultured at 37 °C, 5% CO₂ for 24 h prior to lentiviral transduction and 48 h prior to RNP electroporation.

CHO-K1 cells expressing human ET_A were maintained as monolayer cultures on 100-mm cell culture dishes in Ham’s F12 medium supplemented with 10% fetal bovine serum (FBS) and 1 × antibiotic–antimycotic solution at 37 °C in a humidified atmosphere containing 5% CO₂. The established human colorectal cancer cell lines HT-29 and HCT-116 were purchased from the Korean Cell Line Bank (Seoul, Korea) and maintained in HyClone RPMI-1640 medium (Cytiva, Marlborough, MA, USA) supplemented with 10% HyClone FBS (Cytiva, Marlborough, MA, USA), 1% penicillin-streptomycin, and 1% sodium pyruvate at 37 °C in a humidified atmosphere of 5% CO₂.