Protocol detail

Culturing and Maintaining Cell Lines

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We purchased the following cell lines from American Type Culture Collection (ATCC): HCT116 (CCL-247), U2OS (HTB-96), HEK293T (CRL-3216), and RPE1 (CRL-4000). Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM), which contained 10% fetal bovine serum (FBS) (General Electric ≥ Healthcare), 100 U/mL penicillin G (Life Technologies), and 100 μg/mL streptomycin (Life Technologies). The patient-driven glioblastoma cell line GBL-67 and neural stem cell NSC-10 were established and maintained (35 (link)) and used in this study following the approval of the Institutional Review Board (IRB) at the Seoul National University Hospital (IRB No. H-1904-117-1028). These cells were cultured in DMEM, which contained 20% FBS (GE Healthcare), 100 U/mL penicillin G (Life Technologies), and 100 μg/mL streptomycin (Life Technologies) in a 5% hypoxia chamber.

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Kwon T., Ra J.S., Lee S., Baek I.J., Khim K.W., Lee E.A., Song E.K., Otarbayev D., Jung W., Park Y.H., Wie M., Bae J., Cheng H., Park J.H., Kim N., Seo Y., Yun S., Kim H.E., Moon H.E., Paek S.H., Park T.J., Park Y.U., Rhee H., Choi J.H., Cho S.W, & Myung K. (2022). Precision targeting tumor cells using cancer-specific InDel mutations with CRISPR-Cas9. Proceedings of the National Academy of Sciences of the United States of America, 119(9), e2103532119.

Publication 2022

HCT116 HEK293T RPE1 penicillin G streptomycin FBS

Cell line Cells Culture cell Cultured medium Eagle Electric Fetal bovine serum Glioblastoma Hypoxia Institutional review board Neural stem cell Patient Penicillin g Streptomycin

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Variable analysis

independent variables

Cell lines: HCT116, U2OS, HEK293T, RPE1, GBL-67, NSC-10

dependent variables

Not explicitly mentioned

control variables

Culture medium: Dulbecco's modified Eagle's medium (DMEM)
Serum: 10% fetal bovine serum (FBS) for HCT116, U2OS, HEK293T, RPE1; 20% FBS for GBL-67, NSC-10
Antibiotics: 100 U/mL penicillin G, 100 μg/mL streptomycin
Hypoxia chamber (5% for GBL-67, NSC-10)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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