Epicm

Primary human retinal pigment epithelial (HRPE) cells (Sciencell, cat: 6540) at passage 5 were used for collecting the conditioned medium (CM). Briefly, HRPE cells were cultured in epithelial cell medium (EpiCM, Sciencell, cat: 4101) containing 10% FBS (Sciencell, cat: 0010), 1% epithelial cell growth supplement (EpiCGS, Sciencell, cat: 4152) and 1% penicillin/streptomycin (P/S, Sciencell, cat: 0503). After reaching 80–90% confluence HRPE cells were starved in EpiCM without any supplements for 12 h, followed by 12 h treatment with/without 50 ng/ml recombinant human PDGF-D protein (R&D, cat: 1159-SB/CF). Cells were rinsed and cultured in the supplement-free Dulbecco’s modified Eagle Medium / Ham’s F-12 Mix (DMEM/F-12) (Corning, cat:10-092-CV) and after 24 h collected medium was filtered through 0.22 μm filter (Millex^TM GP Filter Unit, Millipore, cat: SLGP033NB), and stored at –80°C until use.

All the primary cells in this study were cultured in EpiCM (4101, Sciencell (San Diego, California, USA)), but we modified the kit instruction with the replacement of 2% FBS instead of 10% FBS to culture all single cell clones including the epithelial clone L-2F7 and fibroblast clone L-G33. As immortalized gene vectors harbored antibiotic resistance genes, hygromycin (60224ES03, Yeasen) and blasticidin (60218ES10, Yeasen (Shanghai, China)) were selected at concentrations of 200 and 10µg/ml, respectively. The gallbladder cancer cell (GBC) lines NOZ, GBC-SD, ZJU-0430, and OCUG1 were authenticated by Short tandem (STR) repeat and tested if mycoplasma-free. The four GBC cell lines were cultured in Dulbecco's Modified Eagle Medium (DMEM) (SH30243.01, Hyclone (Logan, Utah, USA)) with 10% FBS and 1% penicillin–streptomycin solution (60162ES76, Yeasen (Shanghai, China)). All the cells were digested by a trypsin solution (40127ES60, Yeasen) before passaging or being frozen with a cell-saving buffer (C30100, NCM (Suzhou, Jiangsu Province, China) Biotech).

A human normal trabecular meshwork (TM) cell line was generously donated by Alcon Laboratories, Texas, USA. A human non‐pigmented ciliary epithelial cell line was kindly supplied by Prof. Miguel Coca Prados, Yale University, MA, USA. A human retinal pigment epithelium (RPE) cell line was a gift from Dr Zeev Dvashi, Kaplan Medical Center, Rehovot, Israel. All cell lines were cultured in high glucose Dulbecco's modified Eagle's medium (DMEM) with 10% FBS (foetal bovine serum), 2 mmol/L L‐glutamine, 100 μg/mL streptomycin and 100 units/mL penicillin (all from Biological Industries, Kibbutz Beit HaEmek, Israel) in a humidified atmosphere of 95% air and 5% CO₂ at 37°C. Primary TM cells were kindly provided by W. Daniel Stamer, Duke Eye Center, Durham, North Carolina, USA. Primary TM cells used in these experiments originated from a 63‐year‐ old individual marked as HTM138.2 (passage 2). Primary TM cells were cultured in low glucose DMEM containing 10% FBS, glutamine and antibiotics. Primary human NPCE was purchased from ScienCell Research Laboratories. Primary NPCE cells were maintained in epithelial cell medium (EpiCM; ScienCell Research Laboratories) supplemented with 2% FBS, 1% epithelial cell growth supplement (EPiCGS; ScienCell Research Laboratories) and 1% penicillin‐streptomycin solution (P/S; ScienCell Research Laboratories).

Aedes albopictus C6/36 cells were grown in 30% RPMI-1640 (Gibco-Life Technologies, Carlsbad, CA, USA), 60% Dulbecco’s modified Eagle’s medium (DMEM, Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco) at 28 °C, as previously described.^{44 (link)} HK2 cells were cultured in DMEM/F12 (1:1, Gibco) supplemented with 10% FBS, 100 IU/mL of penicillin and 100 μg/mL of streptomycin and were maintained at 37 °C in a fully humidified atmosphere with 5% CO₂. Primary hRPTEpiCs were purchased from ScienCell (Carlsbad, CA, USA) and maintained on polylysine (ScienCell)-coated plates in Epithelial Cell Medium (EpiCM, ScienCell) supplemented with 2% FBS (ScienCell), 1% epithelial cell growth supplement (EpiCGS, ScienCell) and 1% penicillin/streptomycin solution (P/S, ScienCell). The SZ01 ZIKV stock^{45 (link)} was kindly provided by Professor Cheng-Feng Qin at the Department of Virology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology. The MR766 ZIKV stock was obtained from ATCC (VR-1838; Manassas, VA, USA).