Transwell Insert, 24-Well, 3.0 µm, PET Membrane,IC-21024-30
Product Introduction
This product is a sterile, ready-to-use Transwell insert system consisting of 24-well cell culture inserts with transparent polyethylene terephthalate (PET) track-etched membranes of 3.0 µm pore size. Transwell inserts provide a permeable support for cell growth, enabling researchers to create a two-compartment culture system with independent access to the apical and basolateral sides of the cell monolayer. The PET membrane allows cells to grow on a porous surface while permitting free diffusion of nutrients, signaling molecules, and pharmacological agents across the membrane barrier. This system is widely used for cell migration and invasion assays, transendothelial electrical resistance measurements, drug transport studies, epithelial and endothelial permeability assessment, and co-culture experiments. The transparent PET membrane enables direct microscopic visualization of cells during and after culture without the need to remove the membrane. Each insert is individually packaged and supplied sterile, ensuring convenience and reproducibility for high-throughput applications.
Product Features
1. Track-Etched PET Membrane: Uniform 3.0 µm pores provide consistent and reproducible results for cell migration, invasion, and permeability assays.
2. Transparent Membrane: Allows direct microscopic observation of cell morphology, monolayer formation, and migration progress without membrane removal.
3. Optimal Pore Density: High pore density facilitates efficient solute and cell transmigration while maintaining membrane mechanical integrity.
4. Tissue Culture Treated: PET membrane surface is optimally treated for cell attachment and growth, supporting a wide variety of adherent cell types.
5. Sterile and Ready-to-Use: Each plate is individually packaged and gamma-sterilized, eliminating the need for pre-treatment or sterilization.
Specifications
Size: 48 inserts per box
Membrane Material: Polyethylene terephthalate (PET)
Membrane Diameter: 6.5 mm
Pore Size: 3.0 µm
Plate Format: 24-well plate
Growth Area: 0.33 cm² per insert
Storage and Stability
Storage Conditions: Store at room temperature (15-25¡ãC) in a dry environment, protected from light and moisture.
Shelf Life: The product is stable for 36 months from the date of manufacture when stored as directed in unopened packaging.
Protocol (For Reference Only)
Important: Use aseptic technique when handling inserts and plates to maintain sterility. Pre-warm culture medium to 37¡ãC before use. Pre-wet the membrane with culture medium before seeding cells to ensure uniform hydration.
1. Plate and Insert Preparation: Open the sterile packaging in a biosafety cabinet, remove the 24-well plate containing inserts, and label appropriately. For cell attachment optimization, pre-coat the PET membrane with extracellular matrix proteins such as fibronectin, collagen, or laminin if required for specific cell types.
2. Cell Seeding for Migration Assays: Harvest and count cells according to standard protocols. Resuspend cells in serum-free or low-serum medium and seed into the upper chamber (insert) at a density of 1¡Á10⁴ to 1¡Á10⁵ cells per insert in 100-200 µL volume. Add 500-750 µL of complete medium containing chemoattractant (e.g., 10% FBS, growth factors, or specific chemokines) to the lower chamber (well).
3. Incubation: Incubate the plate at 37¡ãC in a humidified CO₂ incubator for the desired migration period, typically 4-24 hours for migration assays or 24-72 hours for invasion assays with extracellular matrix coating.
4. Quantification of Migrated Cells: After incubation, carefully remove the insert from the well. Remove non-migrated cells from the upper surface of the membrane using a cotton swab moistened with PBS. Fix migrated cells on the lower surface with 4% paraformaldehyde or 100% methanol for 10-15 minutes. Stain cells with 0.1% crystal violet solution for 10-20 minutes, rinse with PBS, and air dry. Quantify migrated cells by counting under a microscope in multiple random fields per insert, or elute the dye with 10% acetic acid and measure absorbance at 560-590 nm.
5. Permeability and Transport Assays: For permeability studies, seed cells on the insert membrane and culture until a confluent monolayer is formed. Replace medium with fresh medium containing the test compound (e.g., fluorescent dextran, drug) in the apical chamber. At designated time points, sample from the basolateral chamber for quantification of transported compound by fluorescence or absorbance measurement. Calculate apparent permeability coefficient (Papp) based on the flux rate, membrane area, and initial compound concentration.
Precautions
1. Handle inserts gently using fine-tipped forceps to avoid damaging the PET membrane; insert damage will compromise assay reproducibility.
2. Avoid touching the membrane surface with pipette tips during cell seeding, medium addition, or sampling to prevent membrane puncture.
3. Pre-wet the membrane with culture medium for 10-15 minutes before cell seeding to ensure uniform hydration and cell distribution across the membrane surface.
4. For invasion assays, coat the upper membrane surface with a thin layer of Matrigel or other extracellular matrix solution and allow to polymerize at 37¡ãC for 1-2 hours before cell seeding.
5. For research use only. Not for use in diagnostic or therapeutic procedures.
FAQ (Simplified)
Q1: What cell density is recommended for seeding?
A1: Optimal seeding density depends on cell type, assay duration, and membrane pore size. Typically, 1¡Á10⁴ to 1¡Á10⁵ cells per insert is recommended for migration assays. Perform a pilot experiment with cell titration to determine the optimal density for your specific cell type and assay endpoint.
Q2: How should I choose the appropriate pore size?
A2: Pore size should be selected based on cell size and assay type. For most epithelial and cancer cell migration assays, 3.0 µm pores are suitable. Larger pores (5.0 or 8.0 µm) are recommended for endothelial cells, fibroblasts, immune cells, or when studying cell invasion through extracellular matrix barriers. Smaller pores (0.4 or 1.0 µm) are appropriate for permeability and transport studies without cell transmigration.
Q3: Can I reuse the Transwell inserts?
A3: No. Inserts are designed for single use only. PET membranes are delicate and cannot be reliably cleaned, sterilized, or reused without compromising membrane integrity and assay reproducibility.
Q4: How do I prevent cells from growing on the underside of the membrane before the assay?
A4: Ensure proper pre-wetting and hydration of the membrane before seeding. If cells migrate prematurely, reduce the initial seeding density or pre-incubate inserts with serum-free medium in both chambers before placing into chemoattractant-containing lower chambers.
Disclaimer
1. For Research Use Only. Not for use in diagnostic or therapeutic procedures.
2. Due to the variable nature of biological research, optimization of cell seeding density, incubation time, and assay conditions is recommended for specific cell types and experimental systems.
3. This warranty is limited to the replacement of the product. The manufacturer assumes no liability for incidental or consequential damages, including loss of samples or data.
4. Wear appropriate protective clothing and gloves when handling this product.
Ordering Information
Catalog Number: IC-21024-30
Product Name: Transwell Insert, 24-Well, 3.0 µm, PET Membrane, 48/Box
Size: 48 inserts per box
Price: CNY ¥1750.00 / USD $175.00 / EUR €210.00 / JPY ¥31500.00
2023 Version
