Atcc htb 38tm

Cancer cell lines: BxPC-3 (pancreas adenocarcinoma, ATCC^® CRL-1687^TM), and HT-29 (colorectal adenocarcinoma, ATCC^® HTB-38^TM) and normal cell line—WI-38 (human lung fibroblasts, ATCC^® CCL-75TM) were obtained from American Type Culture Collection (ATCC, Rockville, MD, USA). BxPC-3 cells were grown in RPMI-1640 medium supplemented with 10% (v/v) FBS and 1% (v/v) of both antibiotics (streptomycin and penicillin). For the HT-29 cell line, RPMI-1640 medium supplemented with 10% (v/v) FBS and 1% (v/v) of both antibiotics and 1% MEM nonessential amino acids was used to ensure proper cell growth. WI-38 cells were grown in MEM medium supplemented with 10% (v/v) FBS, L-Glutamine, 25 mM Hepes, and 1% penicillin–streptomycin. MycoBlue^TM Mycoplasma Detector kit (Vazyme Biotech, Nanjing, China) was used at least every month for the control of mycoplasma contamination in the cell cultures.
Cells were grown at 37 °C in a humidified atmosphere of 5% CO₂ in the air. The culture medium was changed every 24–48 h. Subculture was performed using 0.25% trypsin/EDTA after cells reached confluence.

The HT-29 human colon adenocarcinoma cell line (ATCC®HTB-38^TM) was purchased from ATCC (Manassas, VA, USA) and maintained in McCoy’s 5 A complete medium (Welgene, Daegu, Korea) supplemented with 10% (v/v) fetal bovine serum (FBS; Hyclone, Logan, UT, USA) and gentamycin (50 g/ml; Sigma-Aldrich, St Louis, MO, USA) at 37 °C in a 5% CO₂-containing humidified atmosphere. Transfections were performed using the Viva Magic transfection reagent (Vivagen, Gyeonggi-Do, Korea) according to the manufacturer’s instructions. HT-29 cells (4.0 × 10⁵ cells/well) were plated to 6-well plates, incubated at 37 °C for 24 h, and then transfected with the various expression vectors. To generate cell lines stably expressing the various versions of syndecan-2, HT-29 cells (1 × 10⁶) were transfected with 1 μg of the indicated expression vectors and then selected for 4 weeks in medium containing 800 μg/ml G418 (EMD Biosciences, San Diego, CA, USA). The surviving clones were individually isolated and analyzed by fluorescence-activated cell sorting (FACS) and reverse transcription polymerase chain reaction (RT-PCR).

Human colorectal cancer cell line HT-29 (ATCC^® HTB-38^TM) and HCT116 (ATCC^® CCL-247^TM) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) by the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan) and maintained in RPMI-1640 (Life Technologies Corp., Carlsbad, CA, USA), supplemented with 10% FBS. The incubation conditions were 5% CO₂ at 37 °C.