Biocoat control inserts

Manufactured by Corning

Sourced in Canada

Biocoat control inserts are a type of lab equipment designed for cell culture applications. They provide a consistent and controlled surface for cell attachment and growth. The inserts are available in various sizes and configurations to accommodate different experimental needs.

Automatically generated - may contain errors

Lab products found in correlation

Biocoat matrigel invasion chamber, by Corning (5 mentions) Fibronectin, by Merck Group (2 mentions) Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions) Synergy ht, by Agilent Technologies (1 mentions) Matrigel invasion chamber, by Corning (1 mentions) Kwik diff stain, by Thermo Fisher Scientific (1 mentions) Recombinant murine m csf, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

BioCoat Control Insert; 8.0 µm; 24-well; BioCoat Control Insert 24-well plate 8.0 μm BioCoat Control Insert‐No ECM BioCoat Control Inserts with 8.0 µm PET Membrane BioCoat Control Cell Culture Inserts BioCoat Control Inserts with 8.0 μm PET membrane Control Inserts

13 protocols using biocoat control inserts

Cell Motility and Invasion Assay

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Infected cells (2.5 × 10⁴) were seeded in serum free medium in Corning^® BioCoat^TM Control Inserts (#354578) or Corning^® GFR Matrigel^® Basement Membrane Matrix Invasion Chambers (#354480) (VWR International, Ontario, Canada) containing growth medium in the bottom chamber. RD and RH30 cells were incubated for 72 and 48 h, respectively. Cells were fixed and stained with the Shandon^TM Kwik-Diff^TM Stain (Thermo Fisher Scientific, Ontario, Canada). Cell motility and invasion were assessed according to manufacturer instructions. Six random fields of view were imaged and analyzed using Northern Eclipse Software (NES, Expix Imaging, Ontario, Canada).

Crawford Parks T.E., Marcellus K.A., Langill J., Ravel-Chapuis A., Michaud J., Cowan K.N., Côté J, & Jasmin B.J. (2017). Novel Roles for Staufen1 in Embryonal and Alveolar Rhabdomyosarcoma via c-myc-dependent and -independent events. Scientific Reports, 7, 42342.

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Matrigel Invasion and Migration Assay

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To assess invasion, 3 × 10⁴–1.5 × 10⁵ cells (depending on the cell line) were plated with serum-free medium into the top chamber of a 24-well BioCoat^TM Matrigel Invasion Chamber with 8 µm pores (Corning Inc., Corning, NY, USA). The bottom chamber was filled with medium containing 10% FBS. Migration was assessed similarly to invasion; however, BioCoat^TM Control Inserts with 8-µm pores (Corning Inc.) were used. Chamber membranes were coated with 10 µg/mL fibronectin (Sigma-Aldrich, St. Louis, MO, USA), especially in assays with U87MG cells. After 24 h in culture, the bottom membranes were fixed in 4% paraformaldehyde and stained with hematoxylin. Cells were quantified in five random fields at 200× magnification.

Higa N., Shinsato Y., Kamil M., Hirano T., Takajo T., Shimokawa M., Minami K., Yamamoto M., Kawahara K., Yonezawa H., Hirano H., Furukawa T., Yoshimoto K, & Arita K. (2019). Formin-like 1 (FMNL1) Is Associated with Glioblastoma Multiforme Mesenchymal Subtype and Independently Predicts Poor Prognosis. International Journal of Molecular Sciences, 20(24), 6355.

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Cell Migration and Invasion Assays

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To assess cell migration in vitro, shPTBP1 and NC lentivirus‐infected MDA‐MB‐231 cells (1 × 10⁴cells in 100 μl DMEM) were separately placed in the top chamber of transwell chambers (8‐μm BioCoat Control Inserts, Corning Costar). The lower chamber was filled with 600 μl DMEM supplemented with 10% FBS. After 24 hr incubation at 37°C, the cells were fixed by 85% alcohol and stained with 0.4% trypan blue. The cells in the top chambers were removed with cotton swabs very carefully and counted (five random fields per well at 100× magnification) under a light microscope. For invasion assay, 5 × 10⁴ cells were plated in the matrigel‐coated chamber and the migration assay was performed.

Wang X., Li Y., Fan Y., Yu X., Mao X, & Jin F. (2018). PTBP1 promotes the growth of breast cancer cells through the PTEN/Akt pathway and autophagy. Journal of Cellular Physiology, 233(11), 8930-8939.

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Transwell Migration and Invasion Assay

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For Transwell migration assay, shVPS35 and Con lentivirus‐infected MDA‐MB‐231 cells (2×10⁴ cells in 100 μl Leibovitz's L-15) were separately placed in the top chamber of transwell chambers (8‐μm BioCoat Control Inserts, Corning Costar). The lower chamber was filled with 600 μl Leibovitz’s L-15 supplemented with 10% FBS. After 24 hours incubation at 37 °C, the cells were fixed and stained. The cells in the top chambers were removed with cotton swabs very carefully and counted (five random fields per well at 100× magnification) under a light microscope. For invasion assay, 3×10⁴ cells were plated in the matrigel‐coated chamber and the migration assay was performed.

Li X., Cao Y., Yu X., Jin F, & Li Y. (2021). A novel autophagy-related genes prognostic risk model and validation of autophagy-related oncogene VPS35 in breast cancer. Cancer Cell International, 21, 265.

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Transwell Migration and Invasion Assays

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For Transwell migration assay, LIFR stable overexpressed UCEC cell line Ishikawa were separately placed in the top chamber of transwell chambers (8‐μm BioCoat Control Inserts, Corning Costar). The lower chamber was filled with 500 μl DMEM supplemented with 10% FBS. After 24 hours incubation at 37°C, the cells were fixed and stained. The cells in the top chambers were removed and counted. For invasion assay, cells were plated in the matrigel‐coated chamber and the migration assay was performed. The cell migration and invasion assays has been described previously in detail (21 (link)).

Zhang F., Wang Y., Li H., Li L., Yang X., You X, & Tang L. (2023). Pan-cancer analysis identifies LIFR as a prognostic and immunological biomarker for uterine corpus endometrial carcinoma. Frontiers in Oncology, 13, 1118906.

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Cell Migration and Invasion Assay Protocol

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BioCoat Control Inserts and BioCoat Matrigel Invasion Chamber (24-well, 8 µm; Corning, Corning, NY, USA) were used for cell migration and invasion assays. Chamber membranes of the control inserts were coated with 10 µg/mL fibronectin (Sigma-Aldrich). Cells were inoculated with serum-free medium into the top chamber, and the bottom chamber was filled with medium containing 10% FBS and 1 ng/mL epidermal growth factor (PeproTech, Rocky Hill, NJ, USA). The numbers of inoculated cells were as follows: MKN45, 3.0 × 10⁵; MKN74, 1.5 × 10⁵; MKN7, 1.5 × 10⁵; and GSU, 1.0 × 10⁵. MKN74 and GSU cells were incubated for 48 h, and MKN45 and MKN7 cells were incubated for 72 h. After incubation, the bottom membranes were fixed using 4% paraformaldehyde and stained with hematoxylin. Cell numbers were counted in six fields under a microscope. These experiments were performed independently at least three times.

Hirano T., Shinsato Y., Tanabe K., Higa N., Kamil M., Kawahara K., Yamamoto M., Minami K., Shimokawa M., Arigami T., Yanagita S., Matushita D., Uenosono Y., Ishigami S., Kijima Y., Maemura K., Kitazono I., Tanimoto A., Furukawa T, & Natsugoe S. (2020). FARP1 boosts CDC42 activity from integrin αvβ5 signaling and correlates with poor prognosis of advanced gastric cancer. Oncogenesis, 9(2), 13.

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Cell Migration and Invasion Assay

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Cells were added to the upper portion of a 24-well chamber and allowed to traverse across a 0.8 μM polycarbonate insert toward a chemoattractant (10% FBS) for 24 h, to analyze migration (Corning BioCoat Control Inserts) or invasion (Corning Matrigel Invasion Chambers). Cells failing to traverse the insert were mechanically removed and remaining cells were methanol-fixed and stained with 1% crystal violet. Wells were destained with 10% acetic acid, 30% methanol. Results were quantified in triplicate using a Synergy HT spectrophotometer (BioTek, 595 nm wavelength).

Hasegawa T., Glavich G.J., Pahuski M., Short A., Semmes O.J., Yang L., Galkin V., Drake R, & Esquela-Kerscher A. (2018). Characterization and Evidence of the miR-888 Cluster as a Novel Cancer Network in Prostate. Molecular cancer research : MCR, 16(4), 669-681.

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Invasion and Migration Assay with Matrigel

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Invasion was assessed using Corning^® BioCoat™ Matrigel^® invasion chambers, according to the manufacturer’s instructions. Cells were collected by treatment with trypsin/EDTA, washed twice with phosphate-buffered saline (PBS), pelleted, and resuspended in medium containing 0.2% fetal bovine serum or 0.1% bovine serum albumin. Cells (5 × 10⁴/ml) were incubated in the presence or absence of 50 μg/ml CS-E (Seikagaku Corporation, Tokyo, Japan) or 10 μM JNK inhibitor SP600125 (Cat. No. 129-56-6, Cayman Chemical Company, Ann Arbor, MI) for 20 min at 25°C, added to the upper chamber, and allowed to invade for 22 h at 37°C in a CO₂ incubator. Complete medium containing 10% fetal bovine serum or 200 ng/ml recombinant human/mouse Wnt-5A (Cat. No. 645-WN/CF) was placed in the bottom well as chemoattractant. Migration was assessed using Corning^® BioCoat™ control inserts according to the manufacturer’s instructions. Cells (5 × 10⁴/ml) were prepared as described above.

Nadanaka S., Tamura J.I, & Kitagawa H. (2022). Chondroitin Sulfates Control Invasiveness of the Basal-Like Breast Cancer Cell Line MDA-MB-231 Through ROR1. Frontiers in Oncology, 12, 914838.

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Cancer Cell Invasion Assay

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Infected cells (2.5 × 10⁴) were seeded in serum free medium in Corning_® BioCoat™ Control Inserts (#354578) or Corning_® GFR Matrigel_® Basement Membrane Matrix Invasion Chambers (#354480) (VWR International, Ontario, Canada) containing growth medium in the bottom chamber. DU145 and PC3 cells were incubated for 24 and 72 h, respectively. Cells were fixed and stained with the Kwik-Diff™ Stain (Thermo Fisher Scientific, Ontario, Canada). Cell motility and invasion were assessed according to manufacturer instructions. Five random fields of view were imaged and analyzed using Northern Eclipse Software (NES, Expix Imaging, Ontario, Canada).

Marcellus K.A., Crawford Parks T.E., Almasi S, & Jasmin B.J. (2021). Distinct roles for the RNA-binding protein Staufen1 in prostate cancer. BMC Cancer, 21, 120.

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Macrophage Migration Towards Intestinal Fibroblasts

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BMDMs were prepared by centrifuging mouse leg bones in RPMI supplemented with 10% FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, and 2 mM L-glutamine. After being filtered through a 70 μm nylon mesh, cells were centrifuged plated in non-treated 10 cm dishes, and incubated for 24 hr. Supernatants containing bone marrow precursors were collected, centrifuged at 1,500 rpm, and cultured in differentiation media (RPMI supplemented with 20% fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin, 2 mM L-glutamine, 0.1 mM nonessential amino acids, 1 mM sodium pyruvate, 0.5 mM β-mercaptoethanol and 50 ng/ml recombinant murine MCSF (Peprotech). 2.5 × 10⁴ BMDMs (7 days in culture) were plated in a transwell chamber (Corning Biocoat control inserts) with 8 μm membrane. BMDMs were allowed to migrate for 8 hours at 37°C, 5% CO₂ in the presence of conditioned medium from intestinal fibroblasts. Cells were fixed in cold methanol and stained with crystal violet. Percentage of area was quantified using ImageJ.

Martinez-Ordoñez A., Duran A., Ruiz-Martinez M., Cid-Diaz T., Zhang X., Han Q., Kinoshita H., Muta Y., Linares J.F., Kasashima H., Nakanishi Y., Omar M., Nishimura S., Avila L., Yashiro M., Maeda K., Pannellini T., Pigazzi A., Inghirami G., Marchionni L., Sigal D., Diaz-Meco M.T, & Moscat J. (2022). Hyaluronan driven by epithelial aPKC deficiency remodels the microenvironment and creates a therapeutic vulnerability in mesenchymal colorectal cancer. Cancer cell, 41(2), 252-271.e9.

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