SubculturingEvery 3-4 days at 8000 cells/cm2Volumes used in this protocol are for 75 cm2flasks; proportionally reduce or increase amount of dissociation solutions for culture vessels of other sizes. Remove and discard spent medium. Briefly rinse with 1X PBS (ATCC 30-2200), 1 mL/25 cm2 ...
Human umbilical vein endothelial cells (HUVECs) were purchased from the American Type Culture Collection (ATCC). These cells were cultured in DMEM including 10 % FBS, 100 μg/mL streptomycin, and 100 U/mL penicillin. It was verified that the cell line was devoid of mycoplasma contamina...
Cell culture Human umbilical vein endothelial cells (HUVECs) were purchased from Lonza (Walkersville, MD, USA) and grown for 2–6 passages in EGM2 culture media (Lonza, Basel, Switzerland) supplemented with 2% fetal bovine serum (FBS). The cultures were maintained in a humidified atmosphere wi...
4.3. Cytotoxicity Assays Cell viability was measured using Cell Counting Assay Kit 8 (CCK-8, Dojindo, kumamoto, Japan) according to the manufacturer's protocol. HUVECs (5 × 103 cells/well) were seeded into 96-well plates in culture medium, maintained in regular growth...
HUVECs were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were grown on 100 × 20 mm2Petri dishes coated with 0.1% gelatin for subculture. Cultures were maintained with endothelial cell medium (Sciencell, Carlsbad, CA, USA) containing 5% fetal bovine ...
Primary hPDMSC isolation and culture hPDMSCs were obtained according to our previous study protocol [22, 23]. The placental villi tissue from the different pregnancy periods was dissected into small pieces and extensively rinsed in PBS with 5% penicillin/streptomycin to remove the blood. After being...
After three times of rinse with PBS, the cell samples were immediately examined under the Nikon fluorescence microscope. 2.5. Immunofluorescence assay After 24 h of treatment with the PM2.5, the cells were processed for immunofluorescence assay according to the standard protocol. Briefly, following ...
The tumor suppressor protein promyelocytic leukemia (PML) is a key regulator of inflammatory responses and tumorigenesis and functions through the assembly of subnuclear structures known as PML nuclear bodies (NBs). The inflammation-related cytokine tumo
Two systems were used, namely, a triple negative MDA-MB-231 breast cancer cell line and a human umbilical endothelial cell line (HUVEC), for the study of autocrine and paracrine BDNF-TrkB loop regulation Full size image Measurement of BDNF levels The level of BDNF in the culture medium was...
Protocol: Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until...