Rsc96

The American Type Culture Collection provided human CRC cell lines SW480, Caco-2, and rat Schwann cell line RSC96 (ATCC, Manassas, VA, USA). All cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen, Carlsbad, CA, USA) containing 10% fetal bovine serum (FBS, Invitrogen, Carlsbad, CA, USA) and 1% penicillin/streptomycin and cultured at 37 °C in humidified air with 5% CO₂. All experiments were performed with mycoplasma-free cells.

Primary SCs were isolated from the sciatic nerve of 1-day-old SD rats and further treated with anti-Thy1.1 antibody (Sigma, St Louis, MO) and rabbit complement (Invitrogen, Carlsbad, CA) to remove the fibroblasts as described previously21 (link). The final cell preparation consisted of 98% SCs, as determined by immunocytochemistry with SC marker anti-S100 (DAKO, Carpinteria, CA). A rat SC line (RSC96) was purchased from the American Type Culture Collection.
Primary SCs and RSC96 SCs were cultured in Dulbecco’s modified eagle medium (DMEM) containing 10% fetal bovine serum (FBS) in a humidified 5% CO₂ incubator at 37 °C. Primary SCs were passaged for no more than 3 times prior to use.
SC cultures were transfected with miR-1 mimic, miR-1 inhibitor, or BDNF siRNA (Ribobio, Guangzhou, China), respectively, using Lipofectamine RNAiMAX transfection reagent (Invitrogen) according to the manufacturer’s instructions.

NIH/3T3 (mouse fibroblast cell line, CRL-1658), 3T3-SA (Swiss albino mouse fibroblast cell line, CCL-92), RSC-96 (rat neuron Schwann cell line, CRL-2765), RAW 264.7 (mouse macrophage cell line, TIB-71), RBL-2H3 (rat basophilic leukemia cell line, CRL-2256), CHO-K1 (Chinese hamster ovary cell line, CCL-61), NCI-H292 (human tumor cell line, CRL-1848), and Caco-2 (human epithelial colorectal adenocarcinoma cell) were purchased from American Type Culture Collection (ATCC) (Rockville, MD, USA).

The Rat Schwann cell line RSC96 was obtained from the American Type Culture Collection (ATCC). The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, United States) supplemented with 10% foetal bovine serum (Gibco, United States) and 1% penicillin/streptomycin (Sigma-Aldrich, United States) in an incubator at 37°C in a humidified 5% CO₂ atmosphere. The cells were cultured in six-well plates at a density of 5 × 10⁵ per well. A normal culture medium was used for the control group. For viral infections, HHV7 was added at a multiplicity of infection (MOI) of 10 until obvious lesions appeared. All the cells were collected after 72 h for subsequent experiments.

The rat Schwann cell line (RSC96) was purchased from the American Type Culture Collection (ATCC), cultured in high-glucose Dulbecco’s modified Eagle’s medium (DMEM, Servicebio), which was supplemented with 10% fetal bovine serum (GIBCO), 100-μg·mL⁻¹ penicillin, and 100-μg·mL⁻¹ streptomycin (Servicebio), and stored in an incubator containing 5% CO₂. Rat embryonic dorsal root ganglion cell lines (F11) were purchased from the European Collection of Authenticated Cell Cultures (ECACC), cultured in L15 medium, supplemented with 10% fetal bovine serum (GIBCO), 100-μg·mL⁻¹ penicillin, and 100-μg·mL⁻¹ streptomycin (Servicebio), and stored in a CO₂-free incubator.