A172 cell mobility was tested by wound healing and transwell assay. The wound was made by the culture insert (ibidi). Cells were cultured in DMEM without serum. Average percentage of area closure was measured 24 hours after wounding by ImageJ. Transwell inserts with 8 μm pores in 24 well plates (Corning, Corelle City, NY, USA) were used for the cell immigration assays. 5 × 104 A172 cells were seeded into the upper chambers with/without various treatments in serum‐free DMEM, while the bottom chambers were filled with 800 μL DMEM with 10% FBS. After incubated for 24 hours, cells were washed with PBS for three times, fixed with 4% paraformaldehyde for 30 minutes, and stained with crystal violet staining solution (Beyotime, Shanghai, China) for 4 hours. Cells on the upper surface were removed and the migrating cells on the lower surface of the inserts were photographed and counted by ImageJ. Cell proliferation was tested by the Cell Counting Kit‐8 according to the manufacturer's instructions. Briefly, cells were cultured in 96 well plates in a concentration of 2 × 104 cells/mL, and optical density (OD) of cells was measured at 450 nm after 24 hours.
In 24 well plates
Manufactured by Corning
Sourced in United States
The 24 well plates are a type of laboratory equipment used to hold and contain samples or solutions for various experimental or analytical purposes. They are designed with 24 individual wells, each capable of holding a small volume of liquid or solid material. The primary function of these plates is to provide a standardized and organized platform for conducting experiments, assays, or other lab-based procedures that require multiple samples or replicates.
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2 protocols using in 24 well plates
1
Assessing A172 Cell Mobility
2
Cell Migration and Invasion Assay
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For the migration assay, when UC-MSCs in 24-well plates (Corning) became confluent, hanging cell inserts with 8-µm pore size permeable membrane (BD Biosciences, San Jose, CA, USA) were put on 24-well plates, and 2.0 × 10 4 MCF-7 cells after serum starvation for 4 h were plated onto inserts and cultured for 18 h. MCF-7 cells remaining on the top side of the inserts were removed, and migratory cells to the bottom side were fixed with methanol (Sigma-Aldrich) and stained with hematoxylin and eosin (Beyotime Biotech). Six random fields per insert were photographed, and migratory cells were counted.
The protocol of the invasion assay was similar to that of the migration assay, and 5.0 × 10 4 MCF-7 cells were plated onto Matrigel (BD Biosciences)-coated inserts with 8-µm pore membrane. After cells were incubated for 24 h, noninvasive cells were removed. The invasive cells were fixed, stained, photographed, and counted.
The protocol of the invasion assay was similar to that of the migration assay, and 5.0 × 10 4 MCF-7 cells were plated onto Matrigel (BD Biosciences)-coated inserts with 8-µm pore membrane. After cells were incubated for 24 h, noninvasive cells were removed. The invasive cells were fixed, stained, photographed, and counted.
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