Tc protector

Peripheral blood samples were obtained from patients using BD Vacutainer™ Blood Collection Tubes (Sodium Heparin/Polyester Gel Samples) (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), who received WT1 peptide pulsed DCs therapy before and after seven sessions of intradermal injections of DC vaccines every two weeks.
PBMCs were collected according to manufacturer’s instructions and cryopreserved using TC protector (DS pharma biomedical, Osaka, Japan) under a temperature condition of −80 to −180 °C.

The E8 fragments of Laminin 211 (LN221F), laminin 411 (LN411F), and laminin 511 (LN511F) were purchased from Nippi, Inc. (Tokyo, Japan). Fibronectin and platelet-poor plasma-derived bovine serum (PDS) were procured from FUJIFILM WAKO (Osaka, Japan). Collagen type IV was purchased from Nitta gelatin Inc. (Osaka, Japan). Human endothelial serum-free medium (HE-SFM), Essential 8 Flex medium, vitronectin (VTN-N), chemically defined lipid concentrate, 1,1′-dioctadecyl-3,3,3′,3′-tetramethyl-indocarbocyanine perchlorate acetylated low-density lipoprotein (Dil-Ac-LDL), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) solution (1 M, pH 7.0–7.6), B-27, Hanks’ balanced salt solution with calcium and magnesium, and lucifer yellow (LY) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Endothelial Cell Basal Medium 2 was procured from Lonza (Basel, Switzerland). Cell culture inserts (1.0 µm transparent PET membrane) for a 12-well plate and Matrigel GFR were purchased from Corning, Inc. (Corning, NY, USA). Fetal bovine serum (FBS), FITC-dextran 4000 (FD4), and gelatin were procured from Sigma-Aldrich Corporation (St. Louis, MO, USA). TC Protector was purchased from DS Pharma Biomedical (Osaka, Japan). KnockOut Serum Replacement (KSR) was procured from Invitrogen (Carlsbad, CA, USA). A-83-01 was purchased from Cayman Chemical (Ann Arbor, MI, USA)