Recombinant ptx3

MCF-7 and MDA-MB-231 human breast cancer cell lines and DU-145 and PC-3 human prostate cancer lines were from American Type Culture Collection (Manassas, VA, USA) and were maintained according to distributors ' instructions. All cells were grown in a 5% CO₂ atmosphere at 37°C. Pyrrolidine dithiocarbamate (PDTC), SP600125, SB203580, PD98059, and anti-β-actin antibody were purchased from Sigma (St. Louis, MO, USA). Recombinant PTX3, IL-lβ, IL-17, IL-23, IL-34 and TNFα proteins and rabbit anti-human PTX3 antibody for ELISA were from R&D Systems (Minneapolis, MN, USA).

This study was approved by the Institutional Review Board of MEDIPOST Co., Ltd. (MP-2015-6-4). Umbilical cord blood (UCB) was collected from the umbilical veins after neonatal delivery with informed maternal consent. Harvested UCB was processed within 24 h of collection. UCB-MSCs, isolated and separated from mononuclear cells with Ficoll-Paque™ PLUS (GE Healthcare, Uppsala, Sweden), were washed or suspended in minimum essential medium-alpha (Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Gibco). The cultures were maintained at 37°C in a humidified atmosphere containing 5% CO₂, and the culture medium was changed twice per week [25 (link)]. For each passage, MSCs were cultured for 5 days, harvested with trypsin-ethylenediaminetetraacetic acid (Gibco), counted, and then reseeded at a cell density of 2,000 cells/cm². We followed the methods of Jin et al. [26 (link)]. In this study, we tested seven UCB-MSC lots that were separated from the UCB samples obtained from different donors; the basic information of these cells is summarized in Supplementary Table 1. In all experiments, UCB-MSCs were used at passage 6. Recombinant PTX3 was obtained from R&D Systems (Minneapolis, MN, USA).