Its premix

L. plantarum ZLP001 was originally isolated in our laboratory, from the ileal mucosa of healthy piglets 4 w after weaning. The strain was identified by the China Center of Industrial Culture Collection (Beijing, China) and is preserved in the China General Microbiological Culture Collection Center (CGMCC No. 7370). L. plantarum ZLP001 cells were cultured in improved de Man, Rogosa, and Sharpe liquid medium (10 g peptone, 5 g yeast powder, 20 g glucose, 10 g beef extract, 5 g sodium acetate, 2 g ammonium citrate dibasic, 2 g dipotassium phosphate, 0.58 g magnesium sulfate, 0.19 g manganese sulfate, 1 mL of Tween-80, and water to 1,000 mL; pH 6.5) at 37°C under anaerobic conditions.
The porcine intestinal epithelial cell line (IPEC-J2) was a generous gift from Dr. Glenn Zhang (Oklahoma State University, Stillwater, OK). The IPEC-J2 cells were cultured in DMEM/F12, a 1 : 1 mixture of Dulbecco's modified Eagle's medium and Ham's F-12 (Gibco™, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco™), streptomycin (100 μg/mL), penicillin (100 U/mL), and 1% ITS premix (5 μg/mL insulin, 5 μg/mL transferrin, 5 ng/mL selenium; ScienCell, San Diego, CA) at 37°C in a 5% CO₂ and 95% air atmosphere with 90% humidity.

A porcine intestinal epithelial cell line, IPEC-J2, was cultured in DMEM/F12, a 1:1 mixture of Dulbecco’s modified Eagle’s medium and Ham’s F-12 (Gibco™, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco™), streptomycin (100 μg/mL), penicillin (100 U/mL), and 1% ITS premix (5 μg/mL insulin, 5 μg/mL transferrin, 5 ng/mL selenium) (ScienCell, San Diego, CA, USA) at 37 °C in an atmosphere of 5% CO₂ and 95% air and 90% humidity. The stable luciferase reporter cell line, IPEC-J2/pBD3-luc, was developed as we described [15 (link)] and maintained in the same medium as IPEC-J2 with additional supplemtion of 1 μg/mL puromycin. A porcine lung alveolar macrophage cell line, 3D4/31 (ATCC CRL-2844), was cultured in Roswell Park Memorial Institute (RPMI 1640, Thermo Fisher Scientific) supplemented with 10% FBS, streptomycin (100 μg/mL), penicillin (100 U/mL), and sodium pyruvate (1 mmol/L) at 37 °C in an atmosphere of 5% CO₂ and 95% air and 90% humidity. Cells were subcultured in complete medium every 3–4 day.

The porcine intestinal epithelial cell line IPEC-J2 was kindly provided by Dr. Glenn Zhang (Oklahoma State University, Stillwater, OK, USA). The cells were cultured in DMEM/F12 complete medium (a 1:1 mixture of Dulbecco’s modified Eagle’s medium and Ham’s F-12; Gibco/Thermo Fisher Scientific, Waltham, MA, USA) containing 10% fetal bovine serum (Gibco), streptomycin (100 μg/mL), penicillin (100 U/mL), and 1% ITS premix (5 μg/mL insulin, 5 μg/mL transferrin, 5 ng/mL selenium) (ScienCell, San Diego, CA, USA) in a humidified incubator at 37 °C in the presence of 5% CO₂. The cells were subcultured every 3–4 days.