Biocoat growth factor reduced matrigel chambers

Manufactured by BD

BD Biocoat Growth Factor Reduced Matrigel™ chambers are three-dimensional (3D) cell culture inserts designed for the assessment of cell migration, invasion, and other cellular behaviors. The chambers are coated with a reduced-growth factor Matrigel matrix, which serves as a barrier for cell migration and invasion studies.

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Lab products found in correlation

Axiovert 200 microscope, by Zeiss (1 mentions) Oligofectamine, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

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3 protocols using biocoat growth factor reduced matrigel chambers

Quantifying Cell Invasion Dynamics

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Cell invasion assays were performed as previously described by us [4 (link)]. Briefly, 5×10⁴ cells were
plated on the top of a BD Biocoat Growth Factor Reduced Matrigel™ chambers with
8μM pores (BD Biosciences) and allowed to invade for 24 hours toward NIH 3T3
conditioned media in the lower chamber. Wells were plated in quadruplicate, with three
wells measured for invasion and one as a loading control for cells in the upper chamber.
After removing cells on the top wells for the three invasion wells, invading cells on the
bottom membrane were fixed with 100% methanol for 5 min, and then stained with
0.5% crystal violet in 20% methanol for 10 min. Wells were washed with
water and dried overnight. Cells were then imaged at 10x using light microscopy and the
number of invading cells counted. Experiments were performed in triplicate and a p-value
of 0.05 or less as measured by a two-sided t-test were determined to be significant.

Pavese J.M, & Bergan R.C. (2014). Circulating tumor cells exhibit a biologically aggressive cancer phenotype accompanied by selective resistance to chemotherapy. Cancer letters, 352(2), 179-186.

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Quantifying Cell Invasion Through Matrigel

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Cell invasion was measured as described by us, with modifications [26] (link). After adding 1×10⁵ cells, in RPMI 1640 with 0.1% BSA, to BD BioCoat Growth Factor Reduced Matrigel chambers with 8 µm membranes, cells invaded for 24 hours toward the lower chamber, containing serum-free NIH 3T3 conditioned media. Assays were conducted at least 3 separate times, each in replicates of N = 3. In some experiments, cells were treated with 10 µM inhibitor 48 hours prior to, and during invasion.
Invading cells, i.e., on the bottom membrane, were methanol fixed, crystal violet stained, and imaged and counted under light microscopy. In parallel wells, cells on the top membrane were not removed, allowing confirmation of equal numbers of total cells loaded. For siRNA experiments, cells were stained using the In Situ β-Galactosidase Staining Kit (Agilent Technologies), followed by imaging and counting invaded and non-invaded β-gal positive cells per well.

Pavese J.M., Ogden I.M., Voll E.A., Huang X., Xu L., Jovanovic B, & Bergan R.C. (2014). Mitogen-Activated Protein Kinase Kinase 4 (MAP2K4) Promotes Human Prostate Cancer Metastasis. PLoS ONE, 9(7), e102289.

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Evaluating miR-218-5p and ARF6 in Cell Invasion

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Capan-1 and Hs 766T were transfected for 72 hours with 100 nM Pre-miR Precursor to hsa-miR-218-5p (Ambion, Inc., Austin, TX), 100 nM Anti-miR miRNA Inhibitor to hsa-miR-218-5p (Ambion, Inc., Austin, TX), or 100 nM FlexiTube siRNA to ARF6 (Qiagen, Valencia, CA) using 9.5 µl of Oligofectamine (Life Technologies, Benicia, CA). After transfection, 5x10 4 Capan-1 cells and 2.5x10 4 Hs766T cells were plated in coated BD BioCoat growth factor-reduced matrigel chambers (BD Biosciences, San Jose, CA) for 24 hours. Cells were plated in serum free media in the top chamber while the bottom chamber contained complete media containing 10% FBS as a chemoattractant.
After 24 hours, the chambers were stained with 50 µl of 0.5% crystal violet in 20% methanol for 1 minute, washed in distilled water, swabbed on the inside to remove noninvasive cells, and allowed to dry overnight. Images of 15 separate fields were acquired using a 40X objective on a Zeiss AxioVert 200 microscope using Axiovision Rel.

Rheinheimer B.A., & Heimark R.L. (2020). mir-218 modulates ARF6 expression and inhibits pancreatic ductal adenocarcinoma invasion.

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