Dulbecco’s Modified Eagle’s Medium (DMEM)

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	supplemented with 10% fetal bovine serum (Welgene) and 1% penicillin/streptomycin (Gibco, Carlsbad, CA, USA) at 37°C in a humidified 5% CO2 incubator.

Protocol tips
supplemented with 10% fetal bovine serum (Welgene) and 1% penicillin/streptomycin (Gibco, Carlsbad, CA, USA) at 37°C in a humidified 5% CO2 incubator.

Publication protocol

HSG cells isolated from human submandibular ducts and A253 cells derived from human squamous carcinoma in submandibular glands were cultured in Dulbecco's Modified Eagle's Medium (Welgene, Daegu, South Korea) supplemented with 10% fetal bovine serum (Welgene) and 1% penicillin/streptomycin (Gibco, Carlsbad, CA, USA) at 37°C in a humidified 5% CO2 incubator. Media change was done three times a week and sub-cultured when cells were approximately 80% confluent. 0.25% trypsin/1.0 mM ethylenediaminetetraacetic acid (Welgene) was used to detach cells from the culture dish. Primary cultured normal human fibroblast cells were obtained from biopsy sample of a patient. Normal tissue was submerged in 70% EtOH for 2 min for the sterilization and washed twice with PBS. Normal tissue was minced in HBSS and transferred to cell culture dish containing DMEM supplemented with 10% FBS and 1% penicillin/streptomycin, and incubated for 7 days. At the day 7 media and unattached cells were completely removed. Attached cells were split into four 100mm dishes and continuously subcultured or cryopreserved. Media was changed twice a week.

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Manufacturer protocol

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