After transient transfection of FH-TET3-pEF or empty vector and treatment of TGF-β1 for 48 h, cells were trypsinized and counted. A total of 1 × 105 cells (for migration assay) or 4 × 105 cells (for invasion assay) in 100 μl serum-free medium was added into millicells (Millipore Co., Bedford, MA, USA) without (for migration assay) or with (for invasion assay) Matrigel (Becton Dickinson Labware, Bedford, MA, USA) coated. 500 μl of medium containing 20 % newborn bovine serum was added into the bottom chambers as the chemotactic factor. After incubation for 24 h (for migration assay) or 48 h (for invasion assay) at 37 °C in 5 % CO2, cells remaining on the upper surface of the filter were removed using cotton swabs. Then the migrated cells were fixed using methyl alcohol and stained using 0.1 % crystal violet. Migratory (or invasive) cells were counted and averaged from images of five random fields (original magnification × 200) captured using an inverted light microscope. The mean values of three duplicate assays were used for statistical analysis.
Millicells
Manufactured by Merck Group
Sourced in United States
Millicells are a type of laboratory equipment used for cell culture applications. They provide a controlled and consistent environment for the growth and maintenance of cells in vitro. Millicells are designed to facilitate the exchange of gases, nutrients, and waste products between the cells and the surrounding culture medium, supporting cell viability and proliferation.
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Lab products found in correlation
10 protocols using millicells
1
TGF-β1-induced Cell Migration and Invasion
2
Cell Migration Assay Protocol
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Cells (1×105/well) in 100 µl of media without serum were added into millicells with 8 µm pore size (Millipore Co., Bedford, MA, USA), inserted into wells containing RPMI 1640 with 20% fetal bovine serum as chemotactic factor. After 24 h incubation, the cells remaining on the upper surface of the filter were removed with a cotton swab, and the migrating cells were fixed with 5% glutaric dialdehyde followed by Giemsa staining for quantitation of cell number. The numbers of migrating cells in three high power fields of the lower surfaces of the membranes were counted. A minimum of three wells were counted per experiment.
3
Cell migration assay protocol
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Cells were trypsinized and counted. A total of 1 × 105 cells in 100 μl serum-free medium were added into millicells (Millipore Co., Bedford, MA, USA). 500 μl of 1640 medium containing 20% newborn bovine serum was added to the bottom chambers as the chemotactic factor. After incubation for 24 at 37 °C. Migratory cells were counted and averaged from images of five random fields (original magnification × 200) captured using an inverted light microscope. Each cell count was performed by three researchers.
4
Cell Migration and Invasion Assay
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After treated, cells were trypsinized and counted. A total of 1×105 cells (for migration assay) or 5×105 cells (for invasion assay) in 100 μl serum-free medium were added into millicells (Millipore Co., Bedford, MA, USA) without (for migration assay) or with (for invasion assay) Matrigel (Becton Dickinson Labware, Bedford, MA, USA) coated. 500 μl of 1640 medium containing 20% newborn bovine serum was added to the bottom chambers as the chemotactic factor. After incubation for 24 h (for migration assay) or 48 h (for invasion assay) at 37°C, cells remaining on the upper surface of the filter were removed using cotton swabs. Then the migrated cells were fixed using methyl alcohol and stained using 0.1% crystal violet. Migratory (or invasive) cells were counted and averaged from images of five random fields (original magnification ×200) captured using an inverted light microscope. The mean values of three duplicate assays were used for statistical analysis.
5
BMP4-Induced 3D Culture of MCF-10A Cells
6
Cell Migration Assay Using Matrigel
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A total of 5 × 105 cells in 100 µl serum-free medium were added into millicells (Millipore Co., Bedford, MA, USA) with Matrigel (Becton Dickinson Labware, Bedford, MA, USA) coated. The cells were cultured in a 5% CO2 incubator at 37℃ for 24 hours, transwell was removed, the cells were carefully cleaned with PBS, fixed with 70% ice ethanol solution for 1 hour, and stained with 0.5% crystal violet dye. Place it at room temperature for 20 minutes, wash it with PBS, wipe the unmigrated cells on the upper side of the room with clean cotton ball, and take photos under the microscope.
7
CaCO2 Monolayer TEER and Permeability Assay
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Twelve‐well plate Millicells (0.4 μm, Millipore Corporation) were used for transepithelial electrical resistance (TEER) assays as described previously. Briefly, 0.5 mL CaCO2 cells at the density of 4 × 105 cells/mL were seeded in the apical chamber that bathed in the basal chamber with 1.0 mL DMEM complete medium for 21 days. Voltage was measured daily using EVOM (WPI), which was multiplied by the area of filter (1.12 cm2) to obtain the TEER in Ohm cm2. DMEM complete medium in apical and basal chamber was refreshed every day. The permeability of FITC‐dextran (Sigma) across the CaCO2 cell monolayer was measured as previously described with modifications. At 21 days, 1.0 mg/mL FITC‐dextran was added on the apical side of monolayers after washed twice with PBS. One millilitre cells in the basal chamber were taken at indicated point, and 1.0 mL pre‐warmed fresh medium was added after each sampling to replenish basal medium. The fluorescence emission at 520 nm was measured with excitation at 490 nm using Synergy H1 microplate reader.
8
Cell Migration Assay Protocol
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Cells were trypsinized and counted. A total of 1×10 5 cells in 100 μl serum-free medium were added into millicells (Millipore Co., Bedford, MA, USA). 500 μl of 1640 medium containing 20 % newborn bovine serum was added to the bottom chambers as the chemotactic factor. After incubation for 24 at 37 °C. Migratory cells were counted and averaged from images of ve random elds (original magni cation ×200) captured using an inverted light microscope. Each cell count was performed by three researchers.
9
Cell Migration and Invasion Assays
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Cells were trypsinized and counted. A total of 1×10 5 cells (for migration assay) or 5×10 5 cells (for invasion assay) in 100 μl serum-free medium were added into millicells (Millipore Co, Bedford, MA, USA) without (for migration assay) or with (for invasion assay) Matrigel (Becton Dickinson Labware, Bedford, MA, USA) coated. 500 μl of 1640 medium containing 20% newborn bovine serum was added to the bottom chambers as the chemotactic factor. After incubation for 24 h (for migration assay) or 48 h (for invasion assay) at 37°C. Migratory (or invasive) cells were counted and averaged from images of five random fields (original magnification ×200) captured using an inverted light microscope. Each cell count was performed by three researchers.
10
Cell Migration and Invasion Assay
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After treated, cells were trypsinized and counted. A total of 1×10 5 cells (for migration assay) or 5×10 5 cells (for invasion assay) in 100 μl serum-free medium were added into millicells (Millipore Co., Bedford, MA, USA) without (for migration assay) or with (for invasion assay) Matrigel (Becton Dickinson Labware, Bedford, MA, USA) coated. 500 μl of 1640 medium containing 20 % newborn bovine serum was added to the bottom chambers as the chemotactic factor. After incubation for 24 h (for migration assay) or 48 h (for invasion assay) at 37 °C, cells remaining on the upper surface of the filter were removed using cotton swabs. Then the migrated cells were fixed using methyl alcohol and stained using 0.1 % crystal violet. Migratory (or invasive) cells were counted and averaged from images of five random fields (original magnification ×200) captured using an inverted light microscope. The mean values of three duplicate assays were used for statistical analysis.
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