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Generation of engineered human iPSCs
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Human iPS cell line hiPSC01 was thawed and cultured for ~5 days in 6-well plates to achieve ~50% confluency. Media was replaced with fresh media containing 5 ug/mL polybrene and lentiviral suspension was added at an approximate multiplicity of infection (MOI) of 30. After 24 h this process was repeated. Following two rounds of transduction, cells were allowed to grow to ~80–85% confluency before being passaged. Cells were expanded and cryopreserved for future use.
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