Matrigel transwell chamber

Manufactured by BD

Sourced in United States

The Matrigel transwell chamber is a laboratory equipment used for cell migration and invasion assays. It consists of a porous membrane coated with Matrigel, a reconstituted basement membrane extract. This setup allows researchers to study the ability of cells to migrate through the Matrigel barrier, which mimics the extracellular matrix.

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Lab products found in correlation

Counting chamber, by Olympus (3 mentions) Matrigel, by BD (2 mentions) Transwell chambers with 8 μm pore, by Corning (1 mentions) Matrigel invasion chamber, by BD (1 mentions) 8 μm transwell inserts, by BD (1 mentions)

Close spelling variants of this product

Matrigel (17210 citations) Transwell chambers (810 citations) Matrigel-coated Transwell chambers (82 citations) Matrigel chambers (47 citations) Matrigel-coated transwell cell culture chambers (10 citations) Transwell Matrigel Invasion Chambers (9 citations) Matrigel pre-coated Transwell chambers (4 citations) Transwell precoated Matrigel chambers (3 citations) Matrigel-coated Boyden transwell chambers (3 citations) Transwell pre-coated matrigel chamber (3 citations)

10 protocols using matrigel transwell chamber

Transwell Assay for Cell Migration and Invasion

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The cell migration assay was performed with Transwell chambers with 8-μm pores (Corning, USA). FLS (1 × 10⁵ cells per chamber) were plated in the upper chambers in duplicate filters. DMEM containing 10% FBS was added to the lower chamber as a chemoattractant. FLS were suspended in serum-free DMEM in the upper chambers. Forty-eight hours after transfection with the miR-27a mimic or miR-27a inhibitor, non-migrating cells were removed from the upper surface, and filters were stained with crystal violet. Migrated cells were counted in five representative microscopic fields (100× magnification).
Cell invasion ability was assessed using Matrigel invasion chambers (BD Biosciences, USA). FLS (5 × 10⁴ cells per chamber) were seeded onto Matrigel Transwell chambers pre-coated with Matrigel (BD Biosciences). The lower chambers were filled with DMEM medium containing 10% FBS, and the FLS in the upper chambers were filled with serum-free DMEM. Forty-eight hours after transfection with the miR-27a mimic or miR-27a inhibitor, cells on the top membrane surface were removed. Cells that penetrated to the bottom were stained with crystal violet and counted, as previously described.

Shi D.L., Shi G.R., Xie J., Du X.Z, & Yang H. (2016). MicroRNA-27a Inhibits Cell Migration and Invasion of Fibroblast-Like Synoviocytes by Targeting Follistatin-Like Protein 1 in Rheumatoid Arthritis. Molecules and Cells, 39(8), 611-618.

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Transwell Assay for Cell Invasion

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Cell invasion was determined using Matrigel Transwell chambers (BD Biosciences, Franklin Lakes, NJ, USA) that were used to evaluate the invasion capacity. The lower chamber was filled with medium supplemented with 20% FBS as the chemotactic agent. After 24-hour incubation, the invading cells were fixed on the membranes, stained with crystal violet, and counted. For the migration test, all operations were consistent with the invasion test, except that the six-well chamber was without Matrigel.

Gao C.L., Sun R., Li D.H, & Gong F. (2018). PIWI-like protein 1 upregulation promotes gastric cancer invasion and metastasis. OncoTargets and therapy, 11, 8783-8789.

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Matrigel Transwell Assay for Cell Invasion

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The invasion of A375 and MV3 cells was analyzed using Matrigel Transwell chambers (8-µm pore size; BD Biosciences). Briefly, the upper Transwell chamber was precoated with 100 µl Matrigel (BD Biosciences) at 37°C for 5 h. Then, 2×10⁵ cells suspended in serum-free RPMI-1640 medium were plated into the upper chamber. The lower chamber was filled with RPMI-1640 medium supplemented with 20% FBS. Following 48 h of incubation, the cells remaining in the upper chamber were removed, while the invasive cells in the lower chamber were fixed with 4% formaldehyde for 15 min at room temperature and stained with 0.1% crystal violet for 20 min at room temperature. The invasive cells were visualized (magnification, ×100) using a light microscope.

Han X., Jia Y., Chen X., Sun C, & Sun J. (2021). lncRNA TINCR attenuates the proliferation and invasion, and enhances the apoptosis of cutaneous malignant melanoma cells by regulating the miR-424-5p/LATS1 axis. Oncology Reports, 46(5), 238.

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Radioresistant Cell Invasion Assay

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Transwell invasion assays were conducted to determine the invasive ability of radioresistant cells, using precoated Matrigel Transwell chambers (BD Bioscience) with 8-µm porous membranes (Corning, Inc.). Briefly, A549 cells were seeded into 24-well plates and then exposed to a 2-Gy dose of radiation. After irradiation, 5x10⁴ A549 cells resuspended in serum-free medium were seeded into the top chamber, and complete medium was added to the lower chamber as a chemoattractant. After 48 h, invasive cells were fixed with 4% paraformaldehyde for 10 min at room temperature, and then stained with 0.1% crystal violet at room temperature for 20 min. The number of invaded cells was counted in five random independent fields per well using a light microscope (Olympus Corporation).

Bai L., Zhang J., Gao D., Liu C., Li W, & Li Q. (2021). Downregulation of high mobility group box 1 enhances the radiosensitivity of non-small cell lung cancer by acting as a crucial target of microRNA-107. Experimental and Therapeutic Medicine, 22(1), 679.

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Evaluating miR-124-3p Regulation of Cell Migration

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To determine the effect of upregulated or downregulated miR-124-3p on cell migratory behavior, a wound-healing assay was performed and the closure of the wounds in each well was evaluated. The two cell lines were seeded onto 35-mm dishes coated with fibronectin, and the cells were allowed to reach 100% confluency. An artificial wound was created on a confluent cell monolayer without FBS using a 200-ml pipette tip 24 h after transfection. The culture medium was refreshed with serum-free medium, and the cells were incubated for additional 24 h at 37°C. Serial images were obtained at 0 and 24 h using a light microscope (Olympus, Tokyo, Japan).
For the invasion assay, 2×10⁴ transfected cells were seeded into the upper chamber of the Matrigel^® Transwell chamber (BD Biosciences, Franklin Lakes, NJ, USA). Medium containing 10% FBS in the lower chamber acted as the chemoattractant. Following incubation for a further 48 h at 37°C, non-invading cells were removed from the upper surface with a cotton swab, and the invaded cells attached to the lower surface of the membrane were fixed with methanol, stained with 0.1% crystal violet and then counted using an Olympus light microscope (Olympus Corporation, Tokyo, Japan).

Zhang L., Chen X., Liu B, & Han J. (2017). MicroRNA-124-3p directly targets PDCD6 to inhibit metastasis in breast cancer. Oncology Letters, 15(1), 984-990.

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Transwell Invasion Assay Protocol

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Matrigel Transwell chamber (BD) full of serum-free medium was for invasion determination. Load of the upper chamber was with 1 × 10⁴ cells. Medium consisting of 20% FBS was employed as a chemical attractant for the lower chamber. After incubation at 37°C for 24 h, fixation of the cells adhered to the lower membrane, stain, count was via a microscope and a counting chamber (Olympus, Tokyo, Japan). Three replicates were presented in each experiment [28 ].

Su P., Mao F., Zhang J., Zhang H., Wang M., Xu Y, & Tian Z. (2021). Circular RNA UBR1 promotes the proliferation, migration, and invasion but represses apoptosis of lung cancer cells via modulating microRNA-545-5p/SSFA2 axis. Bioengineered, 12(2), 12135-12147.

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Transwell Invasion and Migration Assay for HCC

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In the HCC cell invasion assay, the inner surface of the insert coated with Matrigel transwell chamber (2 mg·mL⁻¹, BD Biosciences) used, and for the migration assay, the inner surface had no Matrigel-coated transwell chamber used. For transwell invasion assay, cells were transfected with mock control, miR-mimic-negative control, miR-inhibitor-negative control, miR-221 mimic, miR-221 inhibitor, siRNA negative control, and AEG-1 siRNA. After the transfection, cells were collected and seeded (2 × 10⁵) in the upper chamber (8 μm membrane size) containing a serum-free medium. Lower chamber wells were filled with medium containing 10% FBS and incubated for 48 h with CO₂ at 37 °C. After the incubation, non-invading cells were removed from the upper surface of the filters by wiping with a cotton swab. Invading cells were migrated to the bottom of the membrane and fixed with methanol for 20 min at 37 °C and stained with 0.1% crystal violet. The invasiveness of HCC cells was determined by counting the cells that migrated to the lower side of the filter using a light microscope at 20× magnification (Carl Zeiss, Jena, Germany). For migration assay, the same procedure of invasion assay was performed without Matrigel-coated membrane inserts.

Kannan M., Jayamohan S., Moorthy R.K., Chabattula S.C., Ganeshan M, & Arockiam A.J. (2019). AEG-1/miR-221 Axis Cooperatively Regulates the Progression of Hepatocellular Carcinoma by Targeting PTEN/PI3K/AKT Signaling Pathway. International Journal of Molecular Sciences, 20(22), 5526.

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Transwell Invasion and Migration Assay of Cervical Cancer

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For transwell assay, we have used two different types of AEG-1 siRNA to validate the oncogenic role of AEG-1 in CC. Mock Control, miR mimic negative control, miR inhibitor negative control, miR-375 mimic, miR-375 Inhibitor, siRNA negative control, AEG-1 siRNA, AEG-1 siRNA 2 and HPV 16,18 E6/E7 siRNAs were transfected into cervical cancer cells and after 24 h incubation, cells were collected and seeded (2 × 10⁵) on the top of the 8 μm transwell inserts (BD Biosciences, Bedford, MA, USA) with serum-free DMEM. For invasion assay, the inner surface of the insert coated with Matrigel transwell chamber (2 mg ml⁻¹, BD Biosciences) was used. DMEM with 10% FBS was added to the bottom of the transwell chamber. After 48 h incubation, non-invading cells were removed from the top of the Matrigel with a cotton swab. Invaded cells that reached the lower surface of the matrigel-coated membrane were fixed with methanol and stained with 0.1% crystal violet. The CC cells invasiveness was measured by counting in five randomly selected fields under a light microscope at 20 X magnification (Carl Zeiss). For the migration assay, the procedure was similar to the transwell invasion assay except that the inner surface of the chamber had no matrigel coating.

Jayamohan S., Kannan M., Moorthy R.K., Rajasekaran N., Jung H.S., Shin Y.K, & Arockiam A.J. (2019). Dysregulation of miR-375/AEG-1 Axis by Human Papillomavirus 16/18-E6/E7 Promotes Cellular Proliferation, Migration, and Invasion in Cervical Cancer. Frontiers in Oncology, 9, 847.

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Matrigel Invasion Assay Protocol

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A Matrigel transwell chamber (BD), filled with the serum‐free medium, was used for the invasion assay. A total of 1 × 10⁴ cells were loaded into the upper chamber. Medium containing 20% FBS was used as the chemoattractant in the lower chamber. After 24 hours of incubation at 37°C, the cells that had adhered to the lower membrane were fixed, stained, and counted using a microscope and a counting chamber (Olympus, Tokyo, Japan). Photos were taken in 100x field of vision.

Ai C., Ma G., Deng Y., Zheng Q., Gen Y., Li W., Li Y., Zu L, & Zhou Q. (2020). Nm23‐H1 inhibits lung cancer bone‐specific metastasis by upregulating miR‐660‐5p targeted SMARCA5. Thoracic Cancer, 11(3), 640-650.

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Investigating Cell Migration and Invasion

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The cells were seeded onto 35-mm dishes that had been coated with fibronectin to carry out the wound-healing migration assay. Once the cells had reached 100% confluence, a scratch (approximately 500 µm long) was created on the confluent monolayer using a sterile p200 pipette tip. The cellular debris was then removed by replacing the medium with fresh serum-free medium. During the subsequent 48-h culture of the cells, we obtained images at 0, 24, and 48 h. We used a Matrigel Transwell chamber (BD) filled with serum-free medium for the invasion assay. A total of 1 x 10 4 cells were filled into the upper chamber. Medium containing 10% FBS served as the chemoattractant in the lower chamber. After incubating the chamber at 37°C for 64 h, the cells that were adhered to the lower membrane were fixed, stained, and counted using a microscope and a counting chamber (Olympus, Tokyo, Japan).

Shen Y., Ye Y.F., Ruan L.W., Bao L., Wu M.W, & Zhou Y. (2017). Inhibition of miR-660-5p expression suppresses tumor development and metastasis in human breast cancer. Genetics and molecular research : GMR, 16(1).

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