The Transwell migration assay was performed as described (21 (link)). A total of 5×104 MGC-803 or GES-1 cells were plated on the top chambers of 8 µm pore size Transwell plates (Corning Incorporated, Corning, NY, USA). In order to perform the Matrigel-coated Transwell invasion assay as described (21 (link)), Matrigel and 8×104 MGC-803 or GES-1 cells were plated on the top chambers of 8 µm pore size Transwell plates (Corning Incorporated, Corning, NY, USA). The migration and invasion assays were conducted for 48 h; all experiments were performed at least three times in triplicate.
Transwell plate
Manufactured by Corning
Sourced in United States, China, United Kingdom
Transwell plates are a type of cell culture insert system used for in vitro studies. They consist of a permeable membrane support that separates the upper and lower compartments of a well, allowing for the study of cell migration, transport, and co-culture experiments.
Automatically generated - may contain errors
Lab products found in correlation
Matrigel, by BD (190 mentions)
Matrigel, by Corning (59 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (33 mentions)
Crystal violet, by Beyotime (27 mentions)
Crystal violet, by Merck Group (25 mentions)
Matrigel, by Merck Group (16 mentions)
Matrigel matrix, by BD (15 mentions)
Crystal violet, by Solarbio (14 mentions)
Inverted microscope, by Olympus (13 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (12 mentions)
Matrigel basement membrane matrix, by BD (12 mentions)
Inverted light microscope, by Olympus (8 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (8 mentions)
Caco 2 cell, by American Type Culture Collection (7 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (7 mentions)
Ccl21, by R&D Systems (6 mentions)
Paraformaldehyde (pfa), by Solarbio (6 mentions)
Ecm gel, by Merck Group (6 mentions)
Cxcl12, by R&D Systems (5 mentions)
Matrigel matrix, by Corning (5 mentions)
1 155 protocols using transwell plate
1
Transwell Migration and Invasion Assays
2
Assessing BDNF-AS-mediated GC Cell Migration
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The migration ability of GC cells after BDNF-AS overexpression or knockdown was evaluated using 24-well Transwell plates (8-μm pore size; Corning, USA) that were not coated with Matrigel (Falcon; BD Biosciences, USA). The invasion assay was performed utilizing 24-well Transwell plates (8-μm pore size; Corning, USA) that were precoated with Matrigel. In brief, 105 cells were added to the upper chamber after resuspension in 0.5 ml serum-free DMEM, and 0.75 ml DMEM supplemented with 10% FBS was added to the lower chamber. After incubation for 48 h at 37 °C, we removed the remaining cells from the upper chamber, and the cells on the lower surface of the membrane were fixed with 4% paraformaldehyde and stained with crystal violet (0.5%). Finally, the cells were photographed and counted in four or five selected fields of view.
3
Murine Osteoblast-Osteoclast Co-culture System
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To determine the involvement of Sema4D signaling in tenofovir mediated effects, we constructed a murine primary osteoblast-osteoclast precursor co-culture system based on previously described protocols [34 (link)]. Briefly, co-culture is performed as follows. Adherent BMCs from 6–8-wk-old female C57BL/6 or A2AKO mice (n = 5 each) were seeded at a density of 1 × 105 cell/cm2 density with osteogenic medium in the bottom chamber of transwell plates (Corning, New York, NY, USA), and atotal of 200,000 non-adherent cells were collected and seeded in α-MEM with 30 ng/mLM-CSF for two days in the top chamber of transwell plates. At day 3 (day 0 of differentiation), 30 ng/mLRANKL was added to cultures in the presence/absence of Sema4D antibody 10 ng/mL alone or in combination with tenofovir or dipyridamole 1 µM each. Cells and supernatant were collected at different time points for western blot analysis (n = 5 each).
4
Caco-2 Cell Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Caco-2 cells were acquired from the American Type Culture Collection (Manassas, VA, USA). The cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal calf serum, 4 mmol/L l -glutamine, 50,000 IU/L penicillin, and 50 mg/L streptomycin. The cells were either seeded in 6-well Transwell® plates (Corning, Inc., Corning, NY, USA) at a density of 2 × 105 cells/well or in 75 mm Transwell® plates (Corning, Inc., Corning, NY, USA) at a density of 18.8 × 105 cells/well, and they were grown for 14 days post-confluence.
5
Transwell migration and invasion assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Migration and invasion assays were performed as described previously49 (link). For the Transwell migration assay, 1 × 104 MDA-MB-231, 1 × 104 BT-549 or 3 × 104 MCF-10A cells were plated on the top chambers of 8 μm pore size Transwell plates (Corning). For the Matrigel-coated Transwell invasion assay, Matrigel and 3 × 104 MDA-MB-231, 3 × 104 BT-549 or 8 × 104 MCF-10A cells were plated on the top chambers of 8 μm pore size Transwell plates (Corning). All experiments were performed at least three times in triplicate.
6
Assessing Matrine's Effects on Cell Migration and Invasion
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the wound-healing assay, 2×105 cells (A549 and H1299) were plated in culture-insert wells (Ibidi GmbH, Martinsried, Germany) with DMEM containing 10% FBS at 37°C for 24 h. The culture-insert was subsequently removed and fresh DMEM containing 0.5 mg/ml matrine or an equal amount of PBS was added. The width of the healing monolayer wound was recorded after 24 h. For the migration assay, 3×104 cells in DMEM were seeded into the upper chambers of Transwell plates (Corning, Inc., Corning, NY, USA). Complete medium containing 10% FBS in the bottom chamber was used as a chemoattractant, and 0.5 mg/ml matrine was added to inhibit cell migration. For the invasion assay, 5×104 cells in DMEM were seeded in the upper chambers of Transwell plates with 10% Matrigel (Corning, Inc.) at 37°C for 6 h. Following, DMEM with 10% FBS was used in the bottom chamber as a chemoattractant and 0.5 mg/ml matrine was added to assess cell invasion. Following 24 h migration or invasion at 37°C, cells on the lower membrane of inserts were fixed in 100% methanol at 25°C for 15 min, stained with crystal violet at 25°C for 10 min and counted using light microscopy (magnification, ×100).
7
Cell Migration and Invasion Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell migration was measured using the Transwell plate (Corning, NY, USA) with a 0.4-μm pore polyester membrane. Briefly, cells were suspended at a density of 1.0 × 106/ml and 200 μl of cell suspension was transferred to the upper chamber. The bottom chamber was filled with 600 μl of complete medium. After 24 h of incubation, cells were immobilized with 100% methanol for 10 min and stained with 4 g/L crystal violet solution for 15 min. For cell invasion assay, the experimental procedures were similar to cell migration assay but an invasion chamber 24-well plate (Corning, NY, USA) was used instead of the Transwell plate. For both assays, migrated cells were counted in five randomly selected areas under a 100× microscope field. All assays were performed in triplicate.
8
Chondrogenic Differentiation of BMSC Pellets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We evaluated the induction of chondrogenic differentiation of DCECM, Mg-BGNs-1/DCECM, and Mg-BGNs-2/DCECM scaffolds in BMSC pellets using a Transwell system. The BMSC pellets were formulated as described in our previous study with 5 × 105 BMSCs. The DCECM, Mg-BGNs-1/DCECM, and Mg-BGNs-2/DCECM scaffolds were transferred to the lower well of the Transwell plate (Corning Inc., Corning, NY, USA). The BMSCs were placed on the upper well of the Transwell plate (Corning, USA). Chondrogenic medium (Cyagen, Santa Clara, CA, USA) was added to the wells and changed every 3 days. After culturing in the well for 21 days, the pellets were harvested for analysis.
9
Differentiation of Primary Human Airway Epithelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HRT18 (human rectal tumor cell line) and 293 T (human embryonic kidney 293 cell line transformed with SV40 large T antigen) were obtained from ATCC (Manassas, VA, USA), and maintained in DMEM with 10% fetal bovine serum and 2% penicillin, streptomycin and Fugizone (Life Technologies Inc). Primary HTBE cells were obtained from LifeLine Cell Technology (Frederick, MD, USA) and cultured in BronchiaLife Complete Medium (BronchiaLife Basal Medium with BronchiaLife B/T LifeFactors, LifeLine Cell Techonology, Frederick, MD, USA). Till 80-90% confluence, cells were lifted with brief trypsin digestion and plated on 12-well Corning Transwell plates (collagen-coated permeable, 0.4 μm, St Louis, MO, USA) at a density of 2 × 105 cells per well. After 48 hrs incubation, cells were switched to differentiation medium (1:1 ratio of BronchiaLife medium and DMEM high-glucose medium (Invitrogen) with the addition of 1.1 mM CaCl2 and 25 nM retinoic acid) and maintained for 3 weeks in differentiation media at an air-liquid interface. All cell lines were tested for mycoplasma contamination using Mycoplasma Detection Kt (Macgene, Beijing, China) and they were mycoplasma-free.
10
Paracrine Effect of hBMMSCs on EC Migration
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For investigating the paracrine effect of hBMMSCs on ECs migration, 3 × 104 hBMMSCs were seeded into upper chamber of 6-well Corning Transwell plates (an 0.4 μm pore size; Corning, USA). Then, 5 × 104 ECs were then seeded in the lower chamber. Following growth to 100% confluence, cells were subjected to single vertical scratches using a 200 µL pipette tip. The upper chamber of the transwell, which were already seeded with hBMMSCs, were washed three times with PBS and then co-cultured with ECs, and images were recorded at 0 and 12 h after scratching. To investigate the effect of hBMMSCs exosomes on ECs migration, ECs were seeded in 6-well plates, with hBMMSCs derived exosomes (25 µg/mL) were added. Images were then recorded at 0 and 12 h after scratching using an optical microscope (Lecia, Germany). The rate of wound closure was calculated as follows: (mean wound width-mean remaining width) / mean wound width × 100%.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!