Parent and claudin-4-expressing mouse fibroblast cell lines (L cells) were kindly provided by Dr. S. Tsukita (Kyoto University, Kyoto, Japan) (11 (link)). Both cell lines were cultured in Dulbecco's Modified Eagle's medium supplemented with 10% fetal bovine serum in a 5% CO2 atmosphere at 37°C. Parent and claudin-4-expressing human sarcoma cell lines (HT1080 cells) (ATCC, Virginia, USA) were cultured in Dulbecco's Modified Eagle's medium supplemented with 10% fetal bovine serum in a 5% CO2 atmosphere at 37°C (21 (link)). African Green Monkey kidney normal cells (Vero cells) (JCRB Cell Bank, Osaka, Japan) were cultured in Dulbecco's Modified Eagle's medium supplemented with 10% fetal bovine serum in a 5% CO2 atmosphere at 37°C.
Vero cells
Manufactured by Japanese Collection of Research Bioresources Cell Bank
Sourced in Japan
Vero cells are a continuous cell line derived from the kidney cells of the African green monkey (Cercopithecus aethiops). They are commonly used in various research applications, including virus propagation, toxicology studies, and vaccine development.
Automatically generated - may contain errors
Lab products found in correlation
Minimum essential medium (mem), by Fujifilm (2 mentions)
Ht1080 cells, by American Type Culture Collection (1 mentions)
Advanced modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Shandon cytospin 4, by Thermo Fisher Scientific (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
Minimum essential medium (mem), by Nacalai Tesque (1 mentions)
Tryptose phosphate broth, by Merck Group (1 mentions)
Leibovitz l 15 medium, by Thermo Fisher Scientific (1 mentions)
Sh sy5y cells, by European Collection of Authenticated Cell Cultures (1 mentions)
Dmem high glucose, by Fujifilm (1 mentions)
Dmem nutrient mixture f 12 ham, by Fujifilm (1 mentions)
Bhk 21 cells, by Japanese Collection of Research Bioresources Cell Bank (1 mentions)
Accumax dissociation solution, by Innovative Cell Technologies (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Penicillin streptomycin, by Fujifilm (1 mentions)
A549 cells, by RIKEN BioResource Center (1 mentions)
8 protocols using vero cells
1
Cell Culture Protocols for Comparative Analysis
2
Mycoplasma-infected Vero Cell Preparation and Immunocytochemistry
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Mycoplasma-infected Vero cells were prepared as follows. Vero cells (JCRB 0111) were obtained from the Japanese Collection of Research Bioresources (JCRB). The cells were cultured with advanced modified Eagle's Medium (Invitrogen, Grand Island, NY, USA) containing 5% fetal calf serum (Invitrogen) at 37°C with 5% CO2 for 2 days. Then, 100–200 μl PPLO medium containing each of the eight Mycoplasma strains was added to 5 ml medium containing Vero cells and cultured for an additional 5 days. Non-infected Vero cells were prepared as negative controls. The cells were collected, suspended in PBS, and pasted onto a slide using a centrifuge (Shandon Cytospin 4; Thermo Fisher Scientific Inc., Waltham, MA, USA). Mycoplasma infection was confirmed by staining with DNA-binding fluorescent dye (Hoechst 33258). The slides were preserved at −80°C.
The Vero cells were fixed with 4% formaldehyde at room temperature for 10 min. After fixation, fluorescence immunocytochemistry was performed using the CSA II system according to the manufacturer's protocol. Following peroxidase and protein blocking, the cells were incubated for 1 h with the PAb (1:1000). Rabbit Link (1:5) was applied for 30 min. After treatment with Amplification Reagent for 15 min, counterstaining was performed with 4',6-Diamidino-2-phenylindole. Slides were observed by LSM 510 META laser microscopy.
The Vero cells were fixed with 4% formaldehyde at room temperature for 10 min. After fixation, fluorescence immunocytochemistry was performed using the CSA II system according to the manufacturer's protocol. Following peroxidase and protein blocking, the cells were incubated for 1 h with the PAb (1:1000). Rabbit Link (1:5) was applied for 30 min. After treatment with Amplification Reagent for 15 min, counterstaining was performed with 4',6-Diamidino-2-phenylindole. Slides were observed by LSM 510 META laser microscopy.
3
Cell Culture Protocols for Vero and C6/36 Cells
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Vero cells (Japanese Collection of Research Bioresources Cell Bank (JCRB) (Ibaraki, Japan), Cat# JCRB9013) were cultured in modified Eagle’s medium (MEM, Nacalai Tesque, Cat# 21442-25), supplemented with 10% fetal bovine serum (FBS) (HyClone, Cat# SH30396), 1× non-essential amino acids solution (Nacalai Tesque, Cat# 06344-56), and 1× penicillin-streptomycin (P/S, Nacalai Tesque, Cat# 09367-34). C6/36 cells (JCRB, Cat# IFO50010) were cultured at 28 °C in Leibovitz L-15 medium (Thermo Fisher Scientific, Cat #11415064) supplemented with 10% FBS, 0.3% tryptose phosphate broth (Sigma-Aldrich, Cat# T8782-500G), and 1× P/S.
4
Cell Culture Protocol for Various Cell Lines
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Vero cells, obtained from the JCRB Cell Bank (JCRB0111), were cultured in an atmosphere containing 5% CO2 at 37 °C in modified Eagles’ medium (MEM; Wako, Osaka, Japan) supplemented with 10% heat-inactivated fetal bovine serum (FBS). SH-SY 5Y cells, obtained from the European Collection of Authenticated Cell Cultures (94030304), were cultured in an atmosphere containing 5% CO2 at 37 °C in Dulbecco’s MEM (DMEM)/Nutrient Mixture F-12 Ham (Wako) supplemented with 10% heat-inactivated FBS. HEK-293T cells, kindly provided by Dr. Matsuura (Osaka University), were cultured in an atmosphere containing 5% CO2 at 37 °C in high-glucose DMEM (Wako) supplemented with 10% heat-inactivated FBS.
5
Virus Isolation from Tick Homogenates
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Attempts to isolate infectious virus from the aforementioned filtrated tick homogenates were made using Vero cells [African green monkey kidney, Japanese Collection of Research Bioresources Cell Bank (JCRB), Osaka, Japan] or BHK-21 cells (Syrian hamster kidney, JCRB) as described by Kobayashi et al. [23 (link)]. In brief, 50 µL of each filtrate of pooled tick homogenate prepared for RNA virome analysis described above was inoculated onto monolayer cells, which were incubated for 1 h at 37 °C and 5% CO2. A fresh culture medium was then added to each well, and the incubation was continued for seven days, followed by two subsequent blind passages under the same conditions. Culture supernatants after the incubation period were analyzed using next-generation sequencers (as described previously) [23 (link),26 (link)] to confirm viral isolation.
6
Vero Cell Culture Protocol
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Vero cells (JCRB9013) were obtained from the Japanese Collection of Research Bioresources (JCRB) Cell Bank. Note that the quality of the cells was assured by standard tests of the cell bank. Cells were cultivated in a normal growth medium consisting of Eagle’s minimal essential medium (MEM; Fujifilm Wako Pure Chemical Co., Osaka, Japan) supplemented with 5% heat-inactivated fetal bovine serum (FBS; Sigma-Aldrich, MO, USA) and penicillin–streptomycin (Fujifilm Wako Pure Chemical Co.), at 37 °C and under an atmosphere of 5% CO2 and 100% humidity. Cells were detached with AccuMax dissociation solution (Innovative Cell Technologies, CA, USA) and passaged at a split ratio of 1:5.
7
Mosquito Cell Culture and Virus Propagation
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Ae. albopictus C6/36 cells (European Collection of Authenticated Cell Culture) were maintained in minimum essential medium (MEM) containing 10% heat-inactivated fetal bovine serum (FBS) and 2% non-essential amino acids at 28°C with 5% CO2. C6/36 cells were used to propagate virus stock prior to mosquito infection experiments. African green monkey kidney Vero cells (Japanese Collection of Research Bioresources Cell Bank) were maintained in MEM containing 10% FBS at 37°C with 5% CO2. Vero cells were generally used in this study for virus quantification by FFA.
8
Culturing 293T, A549, and Vero Cells
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293T cells were propagated in high-glucose Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum. A549 cells (RIKEN BRC, Tsukuba, Japan) and Vero cells (JCRB Cell Bank, Osaka, Japan) were propagated in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum.
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