Cell invasion assay was carried out with Bio-Coat Matrigel invasion assay system (BD, Bioscience) as recommended in user’s manual instructions. For this, MDA-MB-231 cells (2 × 105 cells/ml) were suspended in DMEM media (serum free) and subsequently plated into the Matrigel transwell chambers consisting of polycarbonate membrane with 8 μm pores. Hereafter, the pre-incubation of cancer cells with or without plant extracts in transwell chambers was conducted for 12 h and then confined chambers were placed specifically into a 24-well plate containing different basal medium. Finally, the cells from upper wells of the transwell chambers were obliterated with autoclaved cotton swab but the cells that shifted to the lower side of the chambers were fixed followed by staining with crystal violet. Invasion ability of the malignant cells was determined by counting the cells in three randomly selected microscopic fields (magnified ×200).
Bio coat matrigel invasion assay system
Manufactured by BD
Sourced in United States
The Bio-Coat Matrigel invasion assay system is a laboratory equipment used to evaluate the invasive potential of cells. It provides a standardized platform for measuring the ability of cells to migrate through a reconstituted basement membrane. The system consists of Matrigel-coated inserts that are placed in a multi-well plate format, allowing for the quantification of invading cells.
Automatically generated - may contain errors
Lab products found in correlation
Dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
Clone 12g5, by R&D Systems (1 mentions)
Clone 11g8, by R&D Systems (1 mentions)
Infinite m200 pro plate reader, by Tecan (1 mentions)
Crystal violet, by Beyotime (1 mentions)
Biorevo, by Keyence (1 mentions)
Thiazolyl blue tetrazolium bromide mtt solution, by Merck Group (1 mentions)
15 protocols using bio coat matrigel invasion assay system
1
Matrigel Invasion Assay of MDA-MB-231 Cells
2
Matrine Inhibits In Vitro Cell Invasion
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In vitro invasion assays were performed with a BD Bio-Coat Matrigel invasion assay system according to the manufacturer's protocol. Cells were seeded 24 h after treatment with different concentrations of matrine for 48 h. Cells suspended in serum-free DMEM-F12 medium (c11330500bt; Invitrogen, Life Technologies) were seeded into the upper chamber, and fetal bovine serum (10%) was added to the bottom chamber. After an incubation for 48 h at 37°C in the presence of 5% CO2, the cells on the upper side were removed with a cotton swab, and the cells on the bottom side of the filter were fixed, stained and counted.
3
Matrigel Invasion Assay for HUVECs
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The in vitro invasion assay was performed using Bio-Coat matrigel invasion assay system (BD Biosciences, San Jose, CA, USA), according to the manufacturer's instructions. 5×104 HUVECs were suspended in Medium 200 and seeded into the Matrigel transwell chambers consisting of polycarbonate membranes with 8 μm pores. After pre-incubation with or without γ-tocotrienol for 12 h, the transwell chambers were then placed into appropriate wells of a 24 well plate, in which either the basal medium only or basal medium containing 10 ng/mL VEGF had been added. After incubation, the upper surfaces of the Transwell chambers were wiped with cotton swabs, and the invading cells were fixed and stained with crystal violet solution. The invading cells were then counted in five randomly selected areas under microscopic observation as described previously [44 (link)].
4
Invasion Assay for CXCR4/CXCR7 Regulation
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The in vitro invasion assay was performed using the Bio-Coat Matrigel invasion assay system (BD Biosciences, San Jose, CA, USA) according to the manufacturer's instructions. Hep3B, Huh7 and Hep3B cells (2×105 cells) were suspended in serum-free medium containing 0.5% BSA and seeded into the Matrigel transwell chambers consisting of polycarbonate membranes with 8-µm pores. After pre-incubation with an anti-CXCR4 antibody (10 µg/ml, Clone 12G5; R&D Systems); an anti-CXCR7 antibody (10 µg/ml, Clone 11G8; R&D Systems); or a specific CXCR4 antagonist, Peptide R (5 µM), which was developed in our laboratory, for 45–60 minutes at 37 °C. Then, CXCL12 or CXCL11 (100 ng/ml) was added to the lower chamber. After 16 h at 37 °C, the cells on the upper surface of the filter were removed using a cotton wool swab. The cells were counted in ten different consecutive high-power fields (magnification: ×200). The anti-CXCR7 monoclonal antibody CXCR7/RDC-1 (Clone 11G8) and the anti-CXCR4 monoclonal antibody against human CXCR4 (Clone 12G5) were purchased from R&D Systems.
5
Invasion Assay of HepG2 Cells
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The in vitro invasion assay was performed using Bio-Coat Matrigel invasion assay system (BD Biosciences, San Jose, CA, USA), according to the manufacturer’s instructions. 1 × 105 HepG2 cells were suspended in serum-free DMEM and seeded into the Matrigel transwell chambers consisting of polycarbonate membranes with 8 μm pores. After pre-incubation with or without BPTT for 8 h, the transwell chambers were then placed into appropriate wells of a 24-well plate, in which either the basal medium only or basal medium containing CXCL12 had been added. After incubation, the upper surfaces of the Transwell chambers were wiped with cotton swabs, and the invading cells were fixed and stained with crystal violet solution. The invading cells were then counted in five randomly selected areas under microscopic observation.
6
Matrigel Invasion Assay for Cell Migration
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Bio-coat matrigel invasion assay system (BD, USA) was used to evaluate cell invasion following the manufacturer’s protocol. Cells were re-suspended in RPMI-1640 medium and seeded into the upper chambers of 24-well transwell plates, and bottom chambers was filled RPMI-1640 medium with FBS (10%). After 24 h incubation, the supernatant cells were removed, while the cells on the bottom were fixed with 4% paraformaldehyde, stained with crystal violet, and counted. All experiments were performed in triplicate.
7
Evaluating Tumor Cell Invasion
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The invasion assay was performed using the Bio-Coat Matrigel invasion assay system (BD Bioscience, USA) following the manufacturer’s protocol. Four groups of SW620 cells were suspended in serum-free RPMI-1640 medium and seeded into the upper chambers of 24-well trans-well plates. FBS (10 %) was added to the bottom chambers. After 24 h, the cells on the upper side were removed with a cotton swab, while the cells on the bottom side of the filter were fixed with 4% paraformaldehyde, stained with crystal violet, and counted. The invasive rate was expressed as a percentage of control. All experiments were performed in triplicates.
8
Cancer Cell Invasion Assay with Pomolic Acid
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In vitro invasion assay was done using Bio-Coat Matrigel invasion assay system (BD Biosciences, Lexington, KY) according to the manufacturer’s instructions. Cancer cells (5 × 104/mL) were suspended in medium and seeded into the Matrigel-precoated transwell chambers with polycarbonate membranes of 8 µm pore size. After preincubation with or without pomolic acid (25 µM), transwell chambers were then placed into 24-well plates, in which was added the basal medium only or basal medium containing 100 ng/mL CXCL12. After incubation (24 hours for MCF7 and MDA-MB-231), the upper surface of transwell chamber was wiped off with a cotton swab and invading cells were fixed and stained with a Diff-Quick stain. The invading cell numbers were counted in 5 randomly selected microscope fields (×100).
9
Evaluating Invasive Potential of C. benghalensis
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The invasive potential of C. benghalensis was gauged by BioCoat Matrigel invasion assay system (BD Biosciences, USA), following the manufacturer's guidelines. The suspension of MDA-MB-231 cells (2×10 5 cells/mL) in serum-free medium was plated in polycarbonate membrane of Matrigel transwell chambers with 8 μ-m pores. Subsequently, preincubated MDA-MB-231 cancer cells (for 12 h) in transwell chambers with or without the extract were placed precisely into a 24-well plate having basal medium. Followed by 12 h incubation, cells at the upper surface were swabbed with cotton while those that invaded through chamber pores were stained with crystal violet. The invaded cell number was counted in three randomly selected images.
10
Matrigel Invasion Assay for Breast Cancer
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The in vitro invasive properties of the breast cancer cells were performed using the BD Bio-Coat Matrigel invasion assay system (BD Biosciences) as previously described [4 (link)]. Briefly, 2 × 105 cancer cells in serum free media were seeded into the Matrigel inserts consisting with 8-um filter pores. After 16 h of incubation, the upper surface of the transwell chambers was removed with a cotton swab and cells that have invaded through the Matrigel were lysed with the Luc-Screen Extended-Glow Luciferase Reporter Gene Assay System kit (Thermo Fisher Scientific, # T1033) and analyzed for luciferase activity using the Tecan Infinite M200 Pro plate reader. In some experiments, invaded cells were counted after being fixed and stained using a crystal violet solution (2% crystal violet, 50% methanol, 10% acetic acid).
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