Culture insert well

Manufactured by Ibidi

Sourced in Germany

Culture-insert wells are specialized laboratory equipment designed for cell culture applications. These wells provide a contained environment for cells to grow and proliferate. The core function of culture-insert wells is to facilitate the controlled culturing of cells in a laboratory setting.

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Lab products found in correlation

Crystal violet, by Merck Group (2 mentions) Formalin, by Merck Group (2 mentions) Pet membrane transwell insert chamber, by BD (2 mentions) Matrigel, by BD (2 mentions) Observer d1 microscope, by Zeiss (1 mentions) Axio observer d1, by Zeiss (1 mentions) Matrigel, by Corning (1 mentions) Transwell plate, by Corning (1 mentions)

Close spelling variants of this product

Culture-Insert (197 citations) Culture-Insert 2 Well (182 citations) Two-well culture inserts (33 citations) Culture-Insert 2 Well in µ-Dish 35 mm (11 citations) Culture-Insert 2 Well in μ-Dish 35 mm (8 citations) Culture-Insert 2 Well system (7 citations) Two-well silicone culture inserts (6 citations) Culture-Insert 4 Well (5 citations) Culture-Insert 3 wells (5 citations) 2-well cell culture inserts (4 citations)

9 protocols using culture insert well

Cancer Cell Migration and Invasion Assays

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For in vitro migration assay, cancer cells (70μL; concentration: 7×10⁵ cells/mL) were added to Culture-Insert well (ibidi) and cultured for 24 hr. After removal of Culture-Insert, cancer cells were cultured for 20 hr. The migration distance of cancer cells was recorded and measured using ImageJ.
For in vitro invasion assay, cancer cells (1.5×10⁵ cells in 200 μL) were suspended in DMEM medium and added to the upper half of a PET membrane transwell insert chamber (BD Biosciences), which was coated with Matrigel (1 mg/mL; BD Biosciences) on a 24-well plate. DMEM medium supplemented with 10% FBS was added as a chemoattractant to the lower half. After incubation at 37°C for 24 hr, cancer cells that passed through the insert were fixed with 3.7% formalin (Sigma-Aldrich) and stained with 0.1% crystal violet (Sigma-Aldrich).

Chou C.K., Fan C.C., Lin P.S., Liao P.Y., Tung J.C., Hsieh C.H., Hung M.C., Chen C.H, & Chang W.C. (2016). Sciellin mediates mesenchymal-to-epithelial transition in colorectal cancer hepatic metastasis. Oncotarget, 7(18), 25742-25754.

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In Vitro Cancer Cell Migration Assay

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The in vitro migration assay was performed using a Culture-Insert well (ibidi). Cancer cells (4.5 × 10⁴ cells) were cultured in suitable media in the device for 24 hr. After removal of the Culture-Insert, cancer cells were cultured for an additional 8 hr. The migration distance of cancer cells was recorded, and the migration area was measured using ImageJ software.

Fan C.C., Cheng W.C., Huang Y.C., Sher Y.P., Liou N.J., Chien Y.C., Lin P.S., Lin P.S., Chen C.H, & Chang W.C. (2017). EFHD2 promotes epithelial-to-mesenchymal transition and correlates with postsurgical recurrence of stage I lung adenocarcinoma. Scientific Reports, 7, 14617.

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Tumor Cell Migration and Invasion Assay

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The wound healing assay was used to study the migration ability of tumor cell in vitro. Tumor cells (70 µL; concentration: 3 × 10⁵ cells/mL) were added to Culture‐Insert well (ibidi) and cultured for 24 h. A “wound gap” was created by removal of Culture‐Insert, and the “healing” of this gap by cell migration was recorded per 4 h until 24 h. The migration area of tumor cells was measured using Image J software. The Matrigel invasion assay was used to determine the invasion ability of tumor cell in vitro. Tumor cells (2.5 × 10⁵ cells in 200 µL) were suspended in DMEM medium and added to the upper half of a PET membrane transwell insert chamber (BD Biosciences), which was coated with Matrigel (1 mg/mL; BD Biosciences) on a 24‐well plate. DMEM medium supplemented with 10% FBS was added as a chemoattractant to the lower half. After incubation at 37°C for 24 h, tumor cells that passed through the insert were fixed with 3% formalin (Sigma‐Aldrich) and stained with 0.2% crystal violet (Sigma‐Aldrich).

Tung J., Huang W., Yang J., Chen G., Fan C., Chien Y., Lin P., Candice Lung S, & Chang W. (2017). Auramine O, an incense smoke ingredient, promotes lung cancer malignancy. Environmental Toxicology, 32(11), 2379-2391.

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Cell Migration Assay with Culture-Insert

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The Culture-Insert Well (ibidi GmbH, Germany) was placed into one well of the 24-well plate using sterile tweezers. Cell suspensions were prepared in full medium, then 8 × 10⁴ cells (in volume 100 µl) were transferred to each of four chambers. The cells were incubated at 37°C and 5% CO₂ to obtain ~100% confluence. After the insert was removed, the cells were gently washed two times with PBS and added to starvation medium. Cell migration was observed by microscope and photographed at intervals (0, 4, 6, 10, 24, 48 h). The area occupied by the cells was determined by TScratch software (18 (link)) as % of open image area. The assay was performed in technical quadruplicate for each cell variant.

Kołat D., Kałuzińska Ż., Bednarek A.K, & Płuciennik E. (2021). Fragile Gene WWOX Guides TFAP2A/TFAP2C-Dependent Actions Against Tumor Progression in Grade II Bladder Cancer. Frontiers in Oncology, 11, 621060.

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Wound Healing Assay with MCF7 Cells and EGF

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The wound-healing assay was performed with Culture-Insert wells (ibidi) coated with 10 µg/mL of collagen. MCF7 cells (3 × 10⁴) were grown as a monolayer for 18 h and then serum-starved for 24 h. A 500 ± 50 µm cell-free gap was generated, and the cells were treated with or without 200 ng/mL EGF. To record the healing process, the closure of the gap was visualized every 5 min for 6 h on a ZEISS observer D1 microscope (Axio Observer D1; Carl Zeiss).

Yang C.Y., Chang P.W., Hsu W.H., Chang H.C., Chen C.L., Lai C.C., Chiu W.T, & Chen H.C. (2019). Src and SHP2 coordinately regulate the dynamics and organization of vimentin filaments during cell migration. Oncogene, 38(21), 4075-4094.

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Assessing Matrine's Effects on Cell Migration and Invasion

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For the wound-healing assay, 2×10⁵ cells (A549 and H1299) were plated in culture-insert wells (Ibidi GmbH, Martinsried, Germany) with DMEM containing 10% FBS at 37°C for 24 h. The culture-insert was subsequently removed and fresh DMEM containing 0.5 mg/ml matrine or an equal amount of PBS was added. The width of the healing monolayer wound was recorded after 24 h. For the migration assay, 3×10⁴ cells in DMEM were seeded into the upper chambers of Transwell plates (Corning, Inc., Corning, NY, USA). Complete medium containing 10% FBS in the bottom chamber was used as a chemoattractant, and 0.5 mg/ml matrine was added to inhibit cell migration. For the invasion assay, 5×10⁴ cells in DMEM were seeded in the upper chambers of Transwell plates with 10% Matrigel (Corning, Inc.) at 37°C for 6 h. Following, DMEM with 10% FBS was used in the bottom chamber as a chemoattractant and 0.5 mg/ml matrine was added to assess cell invasion. Following 24 h migration or invasion at 37°C, cells on the lower membrane of inserts were fixed in 100% methanol at 25°C for 15 min, stained with crystal violet at 25°C for 10 min and counted using light microscopy (magnification, ×100).

Xie W., Lu J., Lu Q., Wang X., Long H., Huang J, & Guo Z. (2018). Matrine inhibits the proliferation and migration of lung cancer cells through regulation of the protein kinase B/glycogen synthase kinase-3β signaling pathways. Experimental and Therapeutic Medicine, 16(2), 723-729.

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Platyphyllenone Wound Healing Assay

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The cells were cultured in culture-insert wells (ibidi, Martinsried, Germany) overnight for self-insertion. After wound formation, the cells were treated with different concentrations of platyphyllenone (2.5, 5, and 10 μM) for 0, 3, 6, and 24 h (pH 7.4). Finally, the photographs of wound closure were obtained, and the mean crawling distance was determined.

Bharath Kumar V., Lin J.T., Mahalakshmi B., Chuang Y.C., Ho H.Y., Lin C.C., Lo Y.S., Hsieh M.J, & Chen M.K. (2021). PlatyphyllenoneExerts Anti-Metastatic Effects on Human Oral Cancer Cells by Modulating Cathepsin L Expression, MAPK Pathway and Epithelial–Mesenchymal Transition. International Journal of Molecular Sciences, 22(9), 5012.

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Wound Healing Assay Using ibidi Culture-Inserts

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Culture-Insert Wells (ibidi, Germany) were used for wound healing assay. The Culture-Inserts were first placed in a 24 well plate. Treated cells were seeded into the separated chambers of the ibidi Culture-Inserts. After cells had grown for approximately 24 h, the Culture-Inserts were removed to create a gap. The process of cell migration was monitored using a microscope at different time points. The distance between the gap was measured. The migration rate of the cells was expressed as relative gap closure.

Cen Y., Zhu T., Zhang Y., Zhao L., Zhu J., Wang L., Xu J., Ding T., Xie X., Wang X, & Lu W. (2021). hsa_circ_0005358 suppresses cervical cancer metastasis by interacting with PTBP1 protein to destabilize CDCP1 mRNA. Molecular Therapy. Nucleic Acids, 27, 227-240.

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Wound Healing Assay with Danshensu

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For self-insertion, appropriate amounts of FaDu and Ca9-22 cells were seeded onto culture-insert wells (Ibidi, Martinsried, Germany) and incubated overnight. Next, the cells were treated with 0, 25, 50, and 100 μM of sodium danshensu for 0, 3, 6, 8, and 24 h after creating the wound. The cells were then photographed and the mean crawling distance was measured.

Kumar V.B., Lin S.H., Mahalakshmi B., Lo Y.S., Lin C.C., Chuang Y.C., Hsieh M.J, & Chen M.K. (2020). Sodium Danshensu Inhibits Oral Cancer Cell Migration and Invasion by Modulating p38 Signaling Pathway. Frontiers in Endocrinology, 11, 568436.

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