Hmec 1 cell line

The HepG2, Caco-2, A549, and 3T3 cell lines (the European Collection of Cell Cultures; ECACC) were obtained from Sigma Chemical Co. (St. Louis, MO, USA), while the HMEC-1 cell line was obtained from ATCC (Salisbury, UK). L-glutamine was obtained from PAA (Pasching, Austria). Cell culture media, such as DMEM and MCDB 131 with supplements, were obtained respectively from GIBCO (Carlsbad, CA, USA) and Cytogen (Sinn, Germany). MitoTracker and YO-PRO 1 were from Invitrogen (Carlsbad, CA, USA). Collagen (Cultrex Rat Collagen I) was obtained from R&D Systems (Minneapolis, MN, USA). All solvents of reagent or analytical grade were purchased from POCH (Gliwice, Poland). All the remaining reagents were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Plastic disposable flasks, 96-well tissue culture plates, pipettes, and tubes were purchased from Nunc (Roskilde, Denmark).

The HMEC-1 cell line (CRL-3243) was purchased from the ATCC cell bank. HMEC-1 cell lyophilization tubes were removed from liquid nitrogen and quickly placed in a 37°C water bath and shaken. After complete thawing, the tubes were centrifuged briefly, sterilized with a 75% alcohol wipe, and transferred to a biosafety cabinet. The supernatant was discarded, and the cells were resuspended and inoculated using a complete medium, gently shaken, and placed in a 5% CO₂ incubator at 37°C. Passaging was performed when the cells reached approximately 80% confluence to maintain good cell growth. HMEC-1 in the logarithmic growth phase was trypsin digested, and 5 × 10⁴ cells/mL cell suspension was made using a complete medium. About 1 mL of cell suspension was inoculated into 12-well culture plates, and the culture was continued to ensure that the spread reached approximately 20% at the time of infection. A 500 uL of infection solution was replaced, and 2.86 uL of virus (7.0E + 8 TU/mL) was added for infection. After 16 h of infection, the fresh culture medium was replaced, and the culture was continued at 37°C. After 72h of infection, the infection efficiency was observed under a fluorescent microscope and photographed. Cells with good growth and successful infection were used for subsequent cytological tests.