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Is considerable cell death expected once NSCs P0 are passaged...

Shivani Goolab Aug 24, 2022

Protocol: Rapid and Simplified Induction of Neural Stem/Progenitor Cells (NSCs/NPCs) and Neurons from Human Induced Pluripotent Stem Cells (hiPSCs)

Is considerable cell death expected once NSCs P0 are passaged and the 5uM ROCKi is removed at day 9? These these cells were seeded at 0.5x10(5) cells/cm2 but they are not confluent within the suggested 4-6 days. Should I seed at 1x10(5) once these cells are confluent?

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Ryutaro Kajihara Author Answered Aug 24, 2022

Department of Biomedical Laboratory Sciences, Faculty of Life Sciences Kumamoto University

In our experience, prolonged treatment with ROCKi do not result in massive cell death, although we recommend ROCKi be removed by replacing fresh medium the next day after cell passages.
Yes, you might want to seed the cells at 1x10(5) cells/cm2 or higher. The higher seeding cell density is, the stabler NSCs are!

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