Pet track etched membrane

Manufactured by Corning

Sourced in United States

The PET track-etched membrane is a filtration product designed for laboratory applications. It is made from polyethylene terephthalate (PET) and features a uniform pore structure created through a track-etching process. The membrane provides precise particle separation and filtration capabilities for various laboratory experiments and analyses.

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Lab products found in correlation

Matrigel, by BD (2 mentions) Biocoat matrigel invasion chamber, by Corning (1 mentions) 4 μm pore pet track etched membranes, by Corning (1 mentions) Transwell unit, by Corning (1 mentions) Fluorescein isothiocyanate labeleddextran fitc dextran, by Merck Group (1 mentions) Knockout serum replacement (ksr), by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Nacalai Tesque (1 mentions)

Spelling variants of this product

PET membrane (16 citations) Track-etched membrane (2 citations) 4-μm pore PET track-etched membranes (2 citations)

7 protocols using pet track etched membrane

Evaluating PLXDC1 Silencing Effects on HUVEC Cell Migration and Invasion

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We performed cell migration and invasion assays to confirm the biological effect on HUVEC cells following PLXDC1 silencing with siRNA. The PLXDC1 siRNA was transfected into the HUVEC cell for 4 h in a CO₂ incubator. After incubation, the HUVEC cells (1 × 10⁵) were collected in EBM-2 serum-free medium, and placed in the upper chamber of a Transwell support coated with 0.1% gelatin (PET track-etched membrane, Corning, NY). The chamber was placed on a 24-well plate, and an EBM-2 medium containing serum was placed in the lower chamber to act as a chemo-attractant. The cells were incubated for 6 h to allow migration. Thereafter, they were stained with 0.1% crystal violet for 30 min, and examined by light microscopy (×400 magnification).
In addition, we performed an invasion assay on HUVECs following PLXDC1 siRNA treatment. Before seeding the cells, we prepared chambers coated with 100 μL of Matrigel (BD Biosciences) at 37 °C for 4 h. HUVEC cells (5 × 10⁴) in serum-free EBM-2 medium were placed in the upper chamber of a Transwell support (PET track-etched membrane, Corning, NY). EBM-2 medium containing serum was then added to the lower chamber. After incubating for 24 h, the lower surface of the membrane was fixed with 4% paraformaldehyde and stained with 0.1% crystal violet. The cells were examined by light microscopy (×400 magnification).

Kim G.H., Won J.E., Byeon Y., Kim M.G., Wi T.I., Lee J.M., Park Y.Y., Lee J.W., Kang T.H., Jung I.D., Shin B.C., Ahn H.J., Lee Y.J., Sood A.K., Han H.D, & Park Y.M. (2018). Selective delivery of PLXDC1 small interfering RNA to endothelial cells for anti-angiogenesis tumor therapy using CD44-targeted chitosan nanoparticles for epithelial ovarian cancer. Drug Delivery, 25(1), 1394-1402.

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Gastric Cancer Cell Migration and Invasion

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The migration assay was performed using transwell 24-well plates with 8-μm pore polyethylene terephthalate (PET) track-etched membranes (CORNING). Matrigel-coated filters (CORNING, BioCoat Matrigel invasion chamber) were used for the invasion assay. Cancer cells were indirectly co-cultured with or without TDMs (ratio of TDMs/GC cells, 2/1) using transwell 6-well plates with 4-μm pore PET track-etched membranes (CORNING) for 48 hours. Then, cells (GCIY: 5 × 10⁴, MKN45: 1 × 10⁵, MKN1: 1 × 10⁴) were plated in the upper chamber with 500 μl MEM or RPMI-1640 containing 0.1% FBS. The lower chamber was filled with MEM or RPMI-1640 containing 15% or 10%FBS, respectively. After incubation for 24 hours at 37°C and 5% CO₂, migrating and invading cells were fixed with 4% paraformaldehyde and stained with 0.5% crystal violet. The number of cells on the lower surface of the membrane in 5 random fields was counted using a bright field light microscope. To examine the effect of IL-6 on migration and invasion ability of GC cells, the same experiment was performed in the medium containing recombinant human IL-6 (R & D, Catalog # 206-IL) at a range of 0–100 ng/ml in the upper chamber (GCIY: 3 × 10⁴, MKN45: 3 × 10⁴) instead of co-culture with TDMs. After interaction for 48 hours, migrating and invading cells were counted in the same way.

Sakamoto S., Kagawa S., Kuwada K., Ito A., Kajioka H., Kakiuchi Y., Watanabe M., Kagawa T., Yoshida R., Kikuchi S., Kuroda S., Tazawa H, & Fujiwara T. (2019). Intraperitoneal cancer-immune microenvironment promotes peritoneal dissemination of gastric cancer. Oncoimmunology, 8(12), e1671760.

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In Vitro Migration and Invasion Assays

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The in vitro migration and invasion assays were performed using 24-well chambers with 8-µm pore size PET track-etched membranes (Corning, Inc., Corning, NY, USA). After transfection for 48 h, a total of 5×10⁴ transfected cells were suspended in the upper chamber with serum-free medium, while the lower chamber was filled with 20% FBS-containing medium. The membranes were covered with Matrigel^® (BD Biosciences, San Jose, CA, USA) to form matrix barriers in the invasion assays. After incubation at 37°C for 24 (migration assay) or 48 h (invasion assay), a cotton swab was used to remove the cells on the upper surface, while the cells on the bottom of the membranes were fixed in 4% methanol solution for 1 h and stained with 0.005% crystal violet for 2 h. These were then counted in three random fields under a fluorescence-inverted microscope (×200 magnification).

Hua Y., Liang C., Miao C., Wang S., Su S., Shao P., Liu B., Bao M., Zhu J., Xu A., Zhang J., Li J, & Wang Z. (2018). MicroRNA-126 inhibits proliferation and metastasis in prostate cancer via regulation of ADAM9. Oncology Letters, 15(6), 9051-9060.

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In Vitro Blood-Brain Barrier Model

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Conventional in vitro BBB modeling was conducted by culturing primary HBMVECs grown in 2D as a flat monolayer by following the published protocols [35 (link)]. Permeability across the monolayer of HBMVECs was measured by using transwell units (24-well format, 0.4 µm pore size with high pore density PET track-etched membrane, cat # 353495), which were made to fit in a 12-well plate, purchased from Corning Incorporated, Corning, NY, USA. To create the BBB model, 1 × 10⁵ cells were seeded onto the transwell inserts and cells were allowed to grow for 2 days in complete classic medium. After endothelial cells become confluent on the transwell, cells were treated in fresh culture medium with 10 µg/mL of either Patient-Exo or Control-Exo for 24 h. The cell monolayer’s ability to limit the penetration of a high molecular weight compound was measured by adding 10 µg/mL fluorescein isothiocyanate-labeled dextran (FITC-dextran; molecular weight: 150 kDa, Sigma-Aldrich, St. Louis, MO, USA) to the culture transwell inserts. In the absence of primary HBMVEC, FITC-dextran added to the culture inserts served as controls. After incubation for 10, 30, 60, and 120 min, 20 µL of medium was collected from the lower compartment, and the fluorescence was measured with a spectrophotometer (BioTek Instruments, Winooski, VT, USA) set to 485/20 nm excitation and 528/20 nm emission.

Chandra P.K., Braun S.E., Maity S., Castorena-Gonzalez J.A., Kim H., Shaffer J.G., Cikic S., Rutkai I., Fan J., Guidry J.J., Worthylake D.K., Li C., Abdel-Mageed A.B, & Busija D.W. (2023). Circulating Plasma Exosomal Proteins of Either SHIV-Infected Rhesus Macaque or HIV-Infected Patient Indicates a Link to Neuropathogenesis. Viruses, 15(3), 794.

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Neurosphere Migration Assay

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Neurospheres were digested with 0.05% trypsin to obtain single neurons and resuspended in Neurobasal medium with 1% B27 Plus Supplement. Cells were seeded on the upper layer of a cell culture insert with PET track-etched membrane (8 μm pore size, 353097, Corning, NY, USA) at density of 1 × 10⁵ cells per well. Neurobasal medium with 2% B27 Plus Supplement was added into the bottom of the lower chamber. After 16 h, the culture insert was taken out and the medium was removed carefully. Cells on the upper surface were wiped, while migrating cells in pore and on the lower surface of membrane were fixed, stained with 0.1% crystal violet and observed.

Li N., Zhou P., Tang H., He L., Fang X., Zhao J., Wang X., Qi Y., Sun C., Lin Y., Qin F., Yang M., Zhang Z., Liao C., Zheng S., Peng X., Xue T., Zhu Q., Li H., Li Y., Liu L., Huang J., Liu L., Peng C., Kaindl A.M., Gecz J., Han D., Liu D., Xu K, & Hu H. (2021). In-depth analysis reveals complex molecular aetiology in a cohort of idiopathic cerebral palsy. Brain, 145(1), 119-141.

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Odontoblast Differentiation from Mouse iPSCs

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Round-bottomed 96-well plates coated with MPC were used to generate EBs from the mouse iPS cells (shown schematically in Fig. 1). After harvesting, the EBs were replaced in 12-well polystyrene plates coated with MPC to grow in suspension with RA for 3 days. Cell aggregates were pooled in non-adherent culture dishes with 1×10 -7 mol/L RA (Wako) for 3 days to form neural crest cells. For differentiation into odontoblast-like cells, the neural crest-induced cells (1.5×10 5 cells/ cm 2 ) were then transferred to a culture system, which consisted of a cell culture insert (10-μm pore size, PET track-etched membrane; Corning, CA, USA) with 10% collagen type-I (Col-I; Nitta gelatin, Osaka, Japan) coated on the upper chamber. This coated upper chamber was then filled with serumfree DMEM (Nacalai Tesque), while the lower chamber was filled with odontoblastic differentiation medium consisting of DMEM (Nacalai Tesque), 15% KSR (Gibco) and recombi-nant 100 ng/ml mouse BMP-4/Carrier-Free (R&D Systems, Minneapolis, MN, USA). After 2 days, these EBs were placed in new Col-Icoated plates and cultured in odontoblastic differentiation medium.

Inoue K., Matsuzaka K, & Inoue T. (2022). Identification of Novel Regulator Involved in Differentiation of Mouse iPS Cells into Odontoblast-like Cells. The Bulletin of Tokyo Dental College, 63(3).

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Transwell Invasion Assay Protocol

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Uninfected or Ctr persistently infected hCEcto, End1/E6E7 and HT1080 cells were detached, counted and added in the upper compartment of a 24-well transwell chamber pre-coated with Matrigel matrix (extracellular matrix) in serum-free medium (SFM).
The growth medium supplemented with 10% FCS, pyruvate and glutamine was placed in the bottom compartment of 24 well as a chemoattractant. Cells were allowed to invade for 24 h at 37 C through extracellular matrix into 8 mm polyethylene terephthalate (PET) track-etched membrane (Corning, Cat# 353097). After incubation, cells in the transwell chamber were fixed in 3.7% paraformaldehyde (PFA), followed by cell permeabilization in 100% methanol and stained with 0.2% crystal violet. The microscopic images were taken before and after swab of Matrigel and data was processed using Adobe Illustrator.

Zadora P.K., Chumduri C., Imami K., Berger H., Mi Y., Selbach M., Meyer T.F, & Gurumurthy R.K. (2019). Integrated Phosphoproteome and Transcriptome Analysis Reveals Chlamydia-Induced Epithelial-to-Mesenchymal Transition in Host Cells. Cell reports, 26(5).

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