Cell migration of HUVECs and MLECs, treated as indicated, was assayed using the Cytoselect™ 24-well Wound Healing Assay kit (Cell Biolabs). Images were captured with a Nikon DSFi1 digital camera coupled to a Nikon ECLIPSE TS100 microscope at × 4 magnification. Cell-free area was quantified with ImageJ software.
Cytoselect 24 well wound healing assay kit
Manufactured by Cell Biolabs
Sourced in United States
The CytoSelect™ 24-Well Wound Healing Assay Kit is a laboratory equipment product designed to measure cell migration and wound healing in vitro. It provides a standardized, reproducible, and high-throughput method for evaluating cellular responses to various treatments or environmental conditions.
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Lab products found in correlation
Eclipse ts100, by Nikon (3 mentions)
Ds fi1, by Nikon (3 mentions)
Qcm ecmatrix cell invasion assay kit, by Merck Group (2 mentions)
Ckx41 bright field microscope, by Olympus (2 mentions)
Crystal violet, by Merck Group (1 mentions)
Dm il led, by Leica (1 mentions)
Hematoxylin, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Vaseline, by Merck Group (1 mentions)
Lucifer yellow, by Merck Group (1 mentions)
Dulbecco s modification of eagle s medium dmem, by Merck Group (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Caffeine, by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Pepsin, by Merck Group (1 mentions)
Formaldehyde, by Merck Group (1 mentions)
Griess reagent, by Merck Group (1 mentions)
Eclipse ts2, by Nikon (1 mentions)
Axio observer 7, by Zeiss (1 mentions)
Cytoselect 24 well cell invasion assay kit, by Cell Biolabs (1 mentions)
22 protocols using cytoselect 24 well wound healing assay kit
1
Wound Healing Assay of Endothelial Cells
2
Cell Migration Assay for Biomaterials
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The migration of cells on the materials (n = 3) was detected using the in vitro CytoSelect™ 24-Well Wound Healing Assay Kit from Cell Biolabs Inc. (San Diego, CA). Briefly, GFP-hBMSCs were seeded at a density of 2 × 105 ml−1 on the different biomaterials through the inserts. After 1 day, the inserts were carefully removed from the wells to generate a 0.9 mm wound gap. The cells on the materials were then allowed to migrate into the gap for 24 and 48 h. After fixation, wound gaps were visualized with a light microscope (Nikon Instruments Europe BV, the Netherlands). The number of cells migrated to the cell-free gap was counted by three blinded observers.
3
Wound Healing Assay Protocol
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Wound healing ability was measured using a CytoSelect 24-Well Wound Healing Assay Kit (Cell Biolabs). The experiment was performed according to the manufacturer’s instructions. SNU387 and SNU398 cells were cultured to full confluence. At the start of the experiment, the inserts were removed and the wells were washed with PBS to removed unattached cells and debris. RPMI 1640 culture medium (500 μl) was added to the wells. Cells were incubated at 37°C to allow the closure of the induced wound. After incubation for 24 to 48 hours, migrated cells were visualized using phase-contrast microscopy.
4
Wound Healing Assay of HUVECs
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A CytoSelect 24‐well wound‐healing assay kit (Cell Biolabs Inc., San Diego, CA, USA) was used according to the manufacturer's instructions. Briefly, 3 × 106 HUVECs were added to each well of the wound‐healing insert and grown to confluence. Then, the inserts were removed from the wells, and the cells were washed twice with PBS. The HUVECs were incubated in the absence or presence of Clone 1 IgG or deglyco C1 IgG (20 μg·mL−1) for 8 h at 37 °C. After staining with crystal violet (Sigma), images were acquired using a light microscope (Leica DM IL LED), and migration distances were measured manually.
5
In Vitro Evaluation of Esophageal Regeneration
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2,2-diphenyl-1-picrylhydrazyl radical (DPPH), 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), potassium persulfate, sodium nitroprusside (SNP), Griess reagent, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride, sodium phosphate, Vaseline, caffeine, hydrochloric acid, pepsin from porcine gastric mucosa, Evans blue (EB), fetal bovine serum (FBS), formaldehyde, hematoxylin, eosin, Lucifer Yellow (LY), Dulbecco’s modification of eagle’s medium (DMEM) and penicillin–streptomycin were obtained by Sigma-Aldrich s.r.l. (Milan, Italy).
All solvents were reagent or HPLC grade and obtained from VWR (Milan, Italy).
REF-FTP78 was supplied by Labomar S.p.A. (Istrana, TV, Italy) as 10 mL stick pack.
CytoSelect™ 24-Well Wound Healing Assay Kit (Catalog Number CBA-120) was provided by Cell Biolabs, Inc. (San Diego, CA, USA).
Swine esophagi from commercially available pigs slaughtered for food purposes were supplied by a local slaughter house (Cosenza, CS, Italy) replacing the slaughter of animals for tissue sampling according to the 3Rs (Reduction, Replacement and Refinement). Therefore, the ethical approval was not needed as no animal had been sacrificed for experimental purposes.
Reconstructed human esophageal epithelium (HO2E/S/5, batch N° 21 HO2E 001) and the maintenance medium (21 SMM 018) were supplied by EpiSkin (Lyon Cedex, France).
All solvents were reagent or HPLC grade and obtained from VWR (Milan, Italy).
REF-FTP78 was supplied by Labomar S.p.A. (Istrana, TV, Italy) as 10 mL stick pack.
CytoSelect™ 24-Well Wound Healing Assay Kit (Catalog Number CBA-120) was provided by Cell Biolabs, Inc. (San Diego, CA, USA).
Swine esophagi from commercially available pigs slaughtered for food purposes were supplied by a local slaughter house (Cosenza, CS, Italy) replacing the slaughter of animals for tissue sampling according to the 3Rs (Reduction, Replacement and Refinement). Therefore, the ethical approval was not needed as no animal had been sacrificed for experimental purposes.
Reconstructed human esophageal epithelium (HO2E/S/5, batch N° 21 HO2E 001) and the maintenance medium (21 SMM 018) were supplied by EpiSkin (Lyon Cedex, France).
6
Cell Migration and Invasion Assays
7
Quantitative Wound Healing Assay
8
Quantitative Wound Healing Assay
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CytoSelect™ 24-Well Wound Healing Assay kit from Cell Biolabs, Inc (Cat# CBA-120) was used according to the manufacturer’s instructions. Wound recovery was determined using ImageJ software to calculate percent migration with the formula (1-(area after 24 h/area at 0 h)) x 100% since the cells were fixed and stained at 0 and 24 h after removal of the inserts. Representative images were taken with an Olympus CKX41 bright field microscope.
9
Quantitative Wound Healing Assay for HUVEC/HAoEC
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7 × 104/well HUVEC or HAoEC transfected with miRNA “mimics” were seeded in tissue culture plates from a “Cytoselect 24-well Wound Healing Assay Kit” (Cell Biolabs) following the manufacturer’s recommendations. Cell migration images were captured with a Nikon DSFi1 digital camera coupled to a Nikon ECLIPSE TS100 microscope at 4× magnification the morning after seeding (0 h) and 24 h later (24 h). The cell-free area was quantified with ImageJ software. The migrated area was calculated by subtracting the value of the non-migrated (cell-free area at time 24 h) from the initial wound area (cell-free area at time 0 h), and then expressing this value as a percentage of wound area at time zero.
When indicated 2 mM (final concentration) hydroxyurea was added to the medium to inhibit cell proliferation during the 24 h incubation period.
When indicated 2 mM (final concentration) hydroxyurea was added to the medium to inhibit cell proliferation during the 24 h incubation period.
10
miR-200b Regulates Wound Healing In Vitro
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Wound healing assay was performed using the ATC ASH-3 and KMH-2 cell lines. The ATC cells were transfected separately with the miR-200b mimic or the scrambled negative control obtained from Ambion (Thermo Fisher Scientific, Inc.). Transfection was performed as aforementioned using Lipofectamine® 2000 and Opti-MEM without FBS. When the cell confluence reached ~80%, wounds were created using a 200-µl pipette tip and were then rinsed with RPMI-1640 medium to remove any free-floating cells and debris using the CytoSelect 24-well wound healing assay kit (Cell Biolabs, Inc). After 4 h, the medium was replaced with fresh RPMI-1640 (with 10% FBS), and after 8 h transfection was repeated, and after 4 h, replacement with RPMI-1640 was repeated. Wound healing was observed at different time points (0, 12, 36 and 72 h) within the scrape line, and representative images were captured using a light microscope at magnifications of ×4 and ×10 (Nikon Eclipse TS2; Nikon Corporation). Nikon NIS-Element BR Analysis v5.11.00 software (Nikon Corporation) was used for the quantification of wound healing assays.
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