Culturing Cell Lines and Organoids

All cell lines were cultured in a humidified incubator at 37° C and 5% C02 in medium supplemented with 10% fetal calf serum (FCS) and Pen/Strep solution (Biological industries). The human colon carcinoma cell line SW480 was maintained in DMEM (Biological industries). The human colon cancer isogenic RKO cell lines were kindly provided by Prof. Bert Vogelstein (The Johns Hopkins University, MD, USA) and maintained in McCoy’s 5A medium supplemented with 2mM L-glutamine. The non-small lung carcinoma cell line, NCI-H1299, was maintained in RPMI-1640. iPSCs were generated as previously described [31 (link)] and maintained on irradiated MEFs in ES medium [DMEM (Biological Industries) containing 15% FCS, 5 mg recombinant human LIF (Millipore), 1 mM glutamine (Biological Industries), 1% nonessential amino acids (Biological Industries), 0.1 mM β-mercaPtoethanol (Invitrogen), 60μg/ml penicillin, and 100 μg/ml streptomycin (Biological Industries)]. Intestinal organoids were generated as previously described [15 (link)]. SK-BR-3 breast cancer cells were maintained in RPMI-1640 medium. Mycoplasma test is being performed routinely.

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Solomon H., Dinowitz N., Pateras L.S., Cooks T., Shetzer Y., Molchadsky A., Charni M., Rabani S., Koifman G., Tarcic O., Porat Z., Kogan-Sakin I., Goldfinger N., Oren M., Harris C.C., Gorgoulis V.G, & Rotter V. (2018). Mutant p53 gain of function underlies high expression levels of colorectal cancer stem cells markers. Oncogene, 37(12), 1669-1684.

Publication 2018

Pen/Strep solution DMEM recombinant human LIF glutamine nonessential amino acids β-mercaPtoethanol penicillin streptomycin

Amino acids Biological Breast cancer Carcinoma Cell line Cells Colon Colon cancer Fetal calf serum Glutamine Human Intestinal Ipscs Lung Mercaptoethanol Mycoplasma Organoids Penicillin Strep Streptomycin

Corresponding Organization :

Other organizations : Weizmann Institute of Science, National and Kapodistrian University of Athens, National Institutes of Health, National Cancer Institute

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Variable analysis

independent variables

Cell line type (SW480, RKO, NCI-H1299, iPSCs, intestinal organoids, SK-BR-3)

dependent variables

Cell growth/proliferation
Cell survival
Cell differentiation
Cell signaling pathways

control variables

Incubator temperature (37°C)
CO2 concentration (5%)
Culture medium (DMEM, McCoy's 5A, RPMI-1640, ES medium)
Serum supplement (10% fetal calf serum)
Antibiotic supplement (Pen/Strep)
Glutamine supplement (2mM L-glutamine)
Other supplements (recombinant human LIF, non-essential amino acids, β-mercaptoethanol)

positive controls

Irradiated MEFs for maintaining iPSCs

negative controls

Mycoplasma testing

Annotations

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