A scratch test is an effective means for the assessment of the migration capacities of NP cells (25 (link)). Following treatment, scratches were introduced vertically on the bottom of the slides, with the fine end of 200 µl tips. The medium was then removed, and the slides were washed with PBS three times to remove the detached cells. Serum-free medium was then added for culturing in an incubator at 37°C, 5% CO2 for 4 h. The slides were observed under a CKX31 microscope (Olympus, Tokyo, Japan), and images were captured under three optimal fields at 0 and 4 h following introduction of the scratch with the tips. Cell migration was analyzed using ImageJ 1.43 software (imagej.nih.gov ). The cell migration distance (µm) was calculated as the scratch width (shown as the distance between the two dotted lines in Fig. 1 ) at 0 h minus the scratch width 4 h following the introduction of scratch injury.
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Scratch Assay for NP Cell Migration
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Other organizations : Nanjing Medical University, Changzhou No.2 People's Hospital, Jiangsu University, Jintan People's Hospital
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Variable analysis
independent variables
- Introduction of scratch injury using the fine end of 200 µl tips
- Cell migration distance (µm), calculated as the scratch width at 0 h minus the scratch width 4 h following the introduction of scratch injury
- Culture conditions: Serum-free medium, incubator at 37°C, 5% CO2 for 4 h
- PBS washes to remove detached cells
- No positive or negative controls were explicitly mentioned in the provided information.
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