Hcmec d3

Human cervical carcinoma cell (HeLa, catalog no. CRM-CCL-2^TM) and mouse mononuclear macrophages (J774A.1, catalog no. TIB-67^TM) were obtained from the American Type Culture Collection (ATCC). The human brain endothelial cell (hCMEC/D3, catalog no. 337728) was purchased from BeNa Culture Collection. The luciferase-transfected glioblastoma cell lines (U251-luc, catalog no. CBP30207L and GL261-luc, catalog no. iCell-0059a) were maintained in our laboratory (Shenzhen Second People’s Hospital, Shenzhen, China). Human astrocytes (catalog no. 1800) and brain vascular pericytes (catalog no. 1200) were purchased from Sciencell. The cells have been confirmed without mycoplasma contamination by mycoplasma detection kit (Beijing Solarbio Science & Technology Co., Ltd., CA1080). All types of cells were cultured in DMEM and supplemented with 10% FBS and 1% (v/v) penicillin-streptomycin. The normoxic cells were incubated in a humidified atmosphere with 5% CO₂ and the hypoxia cells were maintained in an incubator with 1% O₂, 5% CO₂, and 94% N₂. Red blood cells (RBC) were obtained from the peripheral blood of mice.

Tetraethyl orthosilicate (TEOS), ferrous chloride (FeCl₃), ferric chloride (FeCl₂), and sodium citrate were purchased from Xilong Chemical Co., Ltd. (Shantou, China). 3-Aminopropyl-trimethoxysilane (APTMS) was purchased from Acros Organics (Belgium, USA).
N-Hydroxy sulfo-succinimide (NHS), 1-(3-(dimethylamino)-propyl)-3-ethylcarbodiimide hydrochloride (EDC), and FITC-labeled Tat (Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg) peptide were provided by Gier Biochemistry Co., Ltd. (Shanghai, China). BCA protein assay kit was purchased from Applygen Technologies Inc. (Beijing, China). External magnetic field was applied by placing an Nd–Fe–B magnet under the culture plates using an immobilized tube apparatus. Human glioma cell lines U251 and human brain capillary endothelial cells hCMEC/D3 (abbreviated as hCMEC) were purchased from BeNa Culture Collection (Beijing, China). All the cells were cultured in RPMI-1640 media with 10 % fetal bovine serum, 10 % penicillin, and 10 % streptomycin at 37 °C in CO₂ incubator.

The immortalized human brain capillary endothelial cell line hCMEC/D3 purchased from BeNa Culture Collection was cultured in endothelial cell medium (ECM) supplemented with 5% fetal bovine serum (FBS), 1% endothelial cell growth supplement and 1% penicillin/streptomycin (P/S). The Madin-Darby canine kidney cell line MDCK was cultured in a Dulbecco’s modified eagle medium (DMEM) supplemented with 10% FBS and 1% P/S. Both cells were incubated at 37℃ with 5% CO2 and used from passage 8–20.