CytoSelect™ 24-Well Cell Co-Culture System
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- 适用
- 哺乳动物
- 应用范围
- Cellular Assay (CA)
- 品牌
- CytoSelect™
- 样品类型
- Cell Samples
- 产品特性
- The CytoSelect™ 24-Well Cell Co-Culture System provides a unique platform to monitor direct contact between two cell types in a single well. First, cells are cultured until they form a monolayer around the insert, creating a defined 8 mm diameter cell-free zone. Once the insert is removed, a second cell type may be seeded into the exposed zone. The kit contains proprietary treated inserts and sufficient reagents for the evaluation of 24 samples. The inserts are compatible with most adherent cell types and experimental conditions.
- 组件
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- 24-well Cell Co-Culture Plate Assembly : Two 24-well plates containing 12 inserts each (see Figure below)
- Calcein AM (500X) : One vial - 50 μL in DMSO 3
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Tumor Necrosis Factor alpha (TNF alpha) ELISA Kit
TNF alpha 适用: 大鼠 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Interleukin 1, beta (IL1B) ELISA KitIL1B 适用: 大鼠 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Tumor Necrosis Factor alpha (TNF alpha) ELISA KitTNF alpha 适用: 小鼠 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
High Mobility Group Box 1 (HMGB1) ELISA KitHMGB1 适用: 人 Colorimetric Sandwich ELISA 62.5 pg/mL - 4000 pg/mL Plasma, Serum
Amyloid beta 1-40 (Abeta 1-40) ELISA KitAbeta 1-40 适用: 人 Colorimetric Sandwich ELISA 7.813 pg/mL - 500 pg/mL Plasma, Serum, Tissue Homogenate
Amyloid beta 1-42 (Abeta 1-42) ELISA KitAbeta 1-42 适用: 人 Colorimetric Sandwich ELISA 4.688 pg/mL - 300 pg/mL Plasma, Serum, Tissue Homogenate
Lipopolysaccharides (LPS) ELISA Kit适用: Various Species Colorimetric Competition ELISA 12.35 ng/mL - 1000 ng/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
High Mobility Group Box 1 (HMGB1) ELISA KitHMGB1 适用: 小鼠 Colorimetric Sandwich ELISA 46.88 pg/mL - 3000 pg/mL Plasma, Serum
Tumor Necrosis Factor alpha (TNF alpha) ELISA KitTNF alpha 适用: 人 AA 77-233 Colorimetric Sandwich ELISA 15.6 pg/mL - 1000 pg/mL Cell Culture Supernatant, Plasma (EDTA), Plasma (citrate), Plasma (heparin), Serum
Vascular Cell Adhesion Molecule 1 (VCAM1) ELISA KitVCAM1 适用: 人 Colorimetric Sandwich ELISA 1.563-100 ng/mL Plasma, Serum, Tissue Homogenate
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- 应用备注
- Optimal working dilution should be determined by the investigator.
- 实验流程
- The CytoSelect™ 24-well Cell Co-Culture System contains two 24-well plates each containing 12 proprietary treated plastic inserts. Calcein AM is also provided for viewing results with fluorescence microscopy.
- 试剂准备
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1X Calcein AM Solution: Just prior to use, prepare a 1X Calcein AM solution by diluting the provided stocks 1:500 in medium. Vortex thoroughly. Protocol (Must be under sterile conditions) Note: It is recommended that all samples be seeded in triplicate. I. Initial Cell Seeding 1. Using sterile forceps, ensure that the inserts have firm contact with the bottom of the plate well. 2. Add 700 μL of culture medium to each well by carefully inserting the pipet tip through one of the top openings of the insert. 6 3. Harvest and resuspend cells in culture medium at 1 - 2 x 10 cells/mL. Note: Cell seeding density is highly cell line dependent, factoring in cell size, spreading and division. Ideally, the desired monolayer confluency should be 80-90 % . 4. For optimal cell dispersion, add 75 μL of cell suspension to each of the three openings at the top of the insert, for a total of 225 μL per well. Take care to avoid bumping and moving the inserts. 5. Incubate cells in a cell culture incubator overnight or until a monolayer forms. 6. Carefully remove the insert from the well. Use sterile forceps to grab and lift the insert slowly from the plate well. 7. Slowly aspirate and discard the media from the wells. Wash wells with media to remove dead cells and debris. Finally, add media to wells to keep cells hydrated. II. Secondary Cell Seeding 6 1. Harvest and resuspend cells in culture medium at 0.1 - 0.3 x 10 cells/mL. Note: Cell seeding density is highly cell line dependent, factoring in cell size, spreading and division. Ideally, the desired monolayer confluency should be 80-90 %. 4 2. Carefully aspirate and discard the media from the wells. Do not allow wells to dry. 3. Slowly add 500 μL of the cell suspension to each well by carefully pipetting down the wall of the well. 4. Transfer the plate to a cell culture incubator for 4-24 hours to allow firm attachment/spreading. Take care to avoid shaking or bumping the plate. 5. Visualize wells under a light microscope. Repeat wash if wells still have debris or unattached cells. (Optional) Calcein AM Fluorometric Labeling 1. Carefully aspirate and discard the media from the wells. Do not allow wells to dry. 2. Slowly add 500 μL of 1X Calcein AM solution to each well by carefully pipetting down the wall of the well. 3. Incubate the plate 30 minutes at 37 °C. 4. Remove the Calcein AM and wash wells 2 times with medium. 5. After the last wash, add enough medium to cover the cells. 6. Monitor the cells microscopically for the presence of the green Calcein (Ex: 485 nm and Em: 515 nm) fluorescence.
- 限制
- 仅限研究用
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- 储存条件
- -20 °C
- 储存方法
- Upon receipt, store the Cell Co-Culture Plates at room temperature and Calcein AM at -20°C.
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- 背景
- The ability to co-culture multiple cell types has enabled novel ways to study cancer, developmental biology, and tissue engineering. Three-dimensional co-culture of endothelial and smooth muscle cells have enabled the structure of a blood vessel to be mimicked. Co-culture experiments of carcinoma and intratumoral stromal cells have played an important role in understanding breast cancer initiation, promotion, and progression. Additionally, feeder cells are commonly co-cultured with stem cells to maintain pluripotency. A common co-culture method utilizes the Boyden Chamber to culture one cell type in the insert with a second cell type in the plate well. However, this approach does not allow for direct cell-cell interaction, recognized as critical in determining cell behavior and producing a more relevant in vivo- like environment.
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