24 well boyden chamber

Manufactured by BD

Sourced in United States

The 24-well Boyden chambers are a laboratory equipment used to study cell migration and invasion. The device consists of a two-compartment system separated by a porous membrane, allowing cells to migrate from one compartment to the other.

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Lab products found in correlation

Matrigel, by BD (2 mentions) Crystal violet, by Merck Group (2 mentions) Matrigel basement membrane matrix, by BD (2 mentions) Infinite m1000 microplate reader, by Tecan (1 mentions) Ckx31, by Olympus (1 mentions) Diff quick kit, by Siemens (1 mentions) Ix51 microscope, by Adobe (1 mentions) Crystal violet, by Sangon (1 mentions) Inverted microscope, by Leica camera (1 mentions) Dmi4000b, by Leica camera (1 mentions) Matrigel coated insert, by BD (1 mentions) 8 μm pore size polycarbonate membranes, by BD (1 mentions)

19 protocols using 24 well boyden chamber

Evaluating HL Cell Migration and Fibroblast Wound Closure

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Migration of HL cells was studied in a 24-well Boyden chamber with 8.0 µm pores (Falcon/Fisher Scientific). 1 × 10⁶ HL cells were transferred into the upper compartment, while crude fibroblast supernatant or medium (with serum, EV depleted) containing 100 µg/ml EVs were placed in the lower compartment. According to the NTA data, a protein concentration of 100 µg/ml corresponds to about 1 × 10⁹ Hodgkin cell-derived EVs/ml. Migrated cells were counted after an incubation time of 26 h.
The scratch assay was performed to monitor the migration of fibroblasts. 1.5 × 10⁵ HDF_n cells/well were seeded in a 24-well Falcon plate. The cell layer was impaired with a scratch and 100 µg/ml HL EVs or medium added. For every condition, cells were seeded in triplicates and two spots per well were monitored with images being recorded in an interval of 15 min for 24 h. For analysis, the scratch width was determined with ImageJ (National Institutes of Health) after 0, 3, 12, and 21 h, wound closure was calculated with the following formula:

% wound closure=1 - (\frac{scratch width t_{x h}}{scratch width t_{0h}}) \times 100 .

Directed migration was studied with a chemotaxis assay performed in Neuroprobe ChemoTx plates. Migration of Calcein AM (MoBiTe)-labeled HDF_n cells toward 40 µg/ml, 150 mg/ml HL EVs or medium (triplicates), was assessed after 2 h via fluorescence detection with an Infinite M1000 microplate reader (Tecan).

Dörsam B., Bösl T., Reiners K.S., Barnert S., Schubert R., Shatnyeva O., Zigrino P., Engert A., Hansen H.P, & von Strandmann E.P. (2018). Hodgkin Lymphoma-Derived Extracellular Vesicles Change the Secretome of Fibroblasts Toward a CAF Phenotype. Frontiers in Immunology, 9, 1358.

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Cell Migration Assay Protocol

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Migration assays were performed using a 24-well Boyden chamber (BD Falcon, Corning-Costar, New York, NY, United States) with an uncoated 8-mm pore size filter. The cells were collected from the culture dishes and washed twice with PBS. Then, the cells were suspended in smooth muscle cell (SMCM) medium (without FBS), and 3 × 10⁴ cells were seeded into the insertion chamber. Subsequently, the cells were cultured in the bottom chamber (containing 0.6 mL of SMCM medium with 10% FBS) at 37 °C (5% CO₂) for 24 h. Cells were stained with 4,6-diamidino-2-phenylindole and counted under a vertical microscope.

Zhou L.L., Jiao Y., Chen H.M., Kang L.H., Yang Q., Li J., Guan M., Zhu G., Liu F.Q., Wang S., Bai X, & Song Y.Q. (2019). Differentially expressed long noncoding RNAs and regulatory mechanism of LINC02407 in human gastric adenocarcinoma. World Journal of Gastroenterology, 25(39), 5973-5990.

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Cell Invasion Assay Using Boyden Chamber

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To test invasion, 2 × 10⁵ cells were seeded into 500 µL of serum‐free medium using a 24‐well Boyden chamber (BD Biosciences, NewJersey, USA) with Matrigel (BD Biosciences). FBS‐containing medium was added into the bottom chamber. After incubation for 24 h, the invaded cells in lower filters were fixed using methanol and stained with crystal violet (Sigma, MO, USA).

Chen Z., He L., Zhao L., Zhang G., Wang Z., Zhu P, & Liu B. (2022). circREEP3 Drives Colorectal Cancer Progression via Activation of FKBP10 Transcription and Restriction of Antitumor Immunity. Advanced Science, 9(13), 2105160.

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Cancer Cell Migration and Invasion Assay

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We used 200 μl of serum-free DMEM to seed (1 × 10⁵) A549, NCI-H1975, and NCI-H292 cells in the 24-well Boyden chamber (BD Biosciences, USA) with an 8 m pore size. The bottom compartment was filled with a solution containing 20% fetal bovine serum in DMEM medium. To perform the migration assay, we cultured the cancer cells in the membrane devoid of an extracellular matrix covering for 6 h. We seeded the cancer cells in the membrane treated with 50 μl of 1:8 diluted Matrigel (BD Biosciences, USA) for the invasion assay. The Boyden chamber was then incubated for roughly 10 h at 37°C with 5% CO₂. The cells that had passed through the bottom chamber’s insert had been preserved, stained, and then viewed under a microscope.

Zhu L., Zeng Q., Wang J., Deng F, & Jin S. (2023). Cathepsin V drives lung cancer progression by shaping the immunosuppressive environment and adhesion molecules cleavage. Aging (Albany NY), 15(23), 13961-13979.

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Quantifying Cell Migration via Transwell Assay

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To measure changes in cell migration, we performed the Transwell migration assay using a 24-well Boyden chamber (BD Biosciences). In brief, 1 × 10⁴ cells were seeded into the upper well in DMEM (without serum), while the lower compartment was filled with 500 µl of RPMI-1640 supplemented with 10% FBS. After incubation at 37°C for 24 h, the cells on the lower surface of the membrane were fixed, stained, and then counted under a light microscope. The average number of migrated cells from five random optical fields (100× magnification) and triplicate filters was determined. The invasion assay was done by the same procedure, except that the inserts of the chambers were coated with Matrigel.

Wang X, & Chen Z. (2016). Knockdown of CUL4B Suppresses the Proliferation and Invasion in Non-Small Cell Lung Cancer Cells. Oncology Research, 24(4), 271-277.

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Transwell Migration and Invasion Assays

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Transwell migration assays were performed in a 24-well Boyden chamber (6.5 mm diameter, 8.0 μm; BD) according to the manufacturer’s instructions as previously described [3 (link)]. In brief, 5 × 10⁴ cells/well in serum-free medium were seeded in the upper chamber, and the lower chamber was filled with complete media. After 24 h, cells in the upper chamber were gently removed. Migrated cells on the lower side of the membrane were fixed with methanol, stained with 0.1% crystal violet, and then counted at 40× magnification in five random fields per well. For the in vitro invasion assay, experiments were performed with Matrigel matrix-coated Transwell chambers.

Zheng L., Liang H., Zhang Q., Shen Z., Sun Y., Zhao X., Gong J., Hou Z., Jiang K., Wang Q., Jin Y, & Yin Y. (2022). circPTEN1, a circular RNA generated from PTEN, suppresses cancer progression through inhibition of TGF-β/Smad signaling. Molecular Cancer, 21, 41.

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Transwell Migration and Invasion Assay

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Transwell migration assays were performed by a 24‐well Boyden chamber (6.5 mm diameter, 8.0 µm; BD) according to the manufacturer’s instructions. In brief, the stably transfected Hela PTEN^−/− cells or H4 cells were re‐suspended in serum‐free media. Approximately 3 × 10⁴ cells/well were seeded on the upper chamber and incubated at 37°C under 5% CO₂ for 24 h, and the lower chamber was filled with complete media containing 20% FBS. After 24 h, the residual cells in the upper chamber were gently removed. Migrated cells on the lower side of the membrane were fixed with methanol and stained with 0.1% crystal violet and counted at 100× magnification at five random fields per well for statistical analysis while images at 200× magnification are chosen as representative images. In the case of in vitro invasion assay, similar experiments were performed by these transwell chambers but coated with the Matrigel matrix.

Zhang Q., Liang H., Zhao X., Zheng L., Li Y., Gong J., Zhu Y., Jin Y, & Yin Y. (2021). PTENε suppresses tumor metastasis through regulation of filopodia formation. The EMBO Journal, 40(10), e105806.

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Evaluating ADAM15 Silencing on Cell Migration

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To assess the effect of siADAM15 on cell migration, a wound-healing assay was performed. Briefly, FLS were seeded at a density of 4x10 3 cells/well in a 96-well plate. After 48 h of siADAM15 transfection, the cells formed a fluent monolayer and were observed under a fluorescent microscope (CKX31; Olympus). A linear scratch was formed using a 10 µl pipette tip 120 h after infection. Wounded monolayers were washed with PBS to remove detached cells and debris. Transwell migration assays were performed using a 24-well Boyden chamber (6.5 mm diameter, 8.0 µm; BD Biosciences, Mountain View, CA, USA) according to the manufacturer's instructions. Photomicrographs of ten random fields were obtained (original magnification, x100), and cells were counted to calculate the average number of cells that had migrated.
For the in vitro invasion assay, similar experiments were performed using inserts coated using a Matrigel basement membrane matrix (BD Biosciences). Briefly, the Matrigel was diluted in serum-free cold media, placed into the upper chambers of a 24-well Transwell and incubated at 37˚C for 1 h. Cells were resuspended with serum-free DMEM media at a density of 5x10 4 cells/well and incubated for 48 h to evaluate cell migration. All experiments were performed in duplicate.

Gao J., Zheng W., Wang L, & Song B. (2015). A disintegrin and metallproteinase 15 knockout decreases migration of fibroblast-like synoviocytes and inflammation in rheumatoid arthritis. Molecular medicine reports, 11(6).

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Evaluating Breast Cancer Cell Invasion and Migration

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Invasion and migration of breast cancer cells were examined using 24-well Boyden chambers (BD Biosciences) with 8 μM inserts with or without precoated Matrigel Basement Membrane Matrix (BD Biosciences), respectively. Breast cancer cells (10⁵ cells per well) were placed on the inserts in the upper chambers and cultured at 37°C in 5% CO2. MDA-MB-231 after 7 h (migration assay) or 16 h (invasion assay), MCF-7 after 30 h (migration assay) or 48 h (invasion assay), cells on the upper surface of the membrane filter were removed. The migrated and invaded cells that crossed the inserts to the lower surface were fixed with 4% formaldehyde, stained with crystal violet (0.005%, sigma), and counted as cells per field of view under microscopy.

Hu P., Chu J., Wu Y., Sun L., Lv X., Zhu Y., Li J., Guo Q., Gong C., Liu B, & Su S. (2015). NBAT1 suppresses breast cancer metastasis by regulating DKK1 via PRC2. Oncotarget, 6(32), 32410-32425.

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Boyden Chamber Cell Migration Assay

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Cell migration was assessed in 24 well Boyden chambers (BD Biosciences, Franklin Lakes, NJ, USA) as described earlier [37] (link). MDA-MB-231 (5×10⁴) cells incubated or not with NGEN or NGENCuB (1, 10 and 100 µM) were seeded on the upper chamber in a DMEM incomplete medium and allowed to migrate for 22 hours at 37°C and 5% CO₂ in a humidified environment. Then, the cells that remained in the upper chamber were removed using a cotton swab. The cells that migrated to the other side of the upper chamber membrane were fixed with methanol and stained with 1% toluidine blue. Cells were counted using Image J software (public domain software) in 5 fields (100× magnification) per well that essentially covered 80% of the well surface. The average number of cells from each of the triplicates represented the average number of cells that migrated in the different groups. Each experiment had triplicate wells for every treatment group and the assays were repeated three times. The mean of all results from controls was considered as 100% of migration.

Filho J.C., Sarria A.L., Becceneri A.B., Fuzer A.M., Batalhão J.R., da Silva C.M., Carlos R.M., Vieira P.C., Fernandes J.B, & Cominetti M.R. (2014). Copper (II) and 2,2′-Bipyridine Complexation Improves Chemopreventive Effects of Naringenin against Breast Tumor Cells. PLoS ONE, 9(9), e107058.

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