Twenty four-well plate cell culture inserts with 8.0 μm pores size (BD BioSciences, Franklin Lakes, NJ, USA) were coated with 50 μl of 0.5 mg/ml Matrigel® matrix solution (BD Biosciences). Serum-containing media without selective antibiotic was placed in the lower chambers of wells, and 200 μl of cell suspension at 5 × 104 cells/ml in serum-free media without selective antibiotics was dispensed onto the matrix coated inserts. Cells were incubated for 72 hours at 37°C in 5% CO2. After removing noninvading cells from upper chamber with a cotton swab, invading cells were fixed with 10% formalin. After inserts were stained with hematoxylin, cells were counted visually by light microscopy. Four inserts were examined for each cell line per experiment. The experiment was repeated three times.
Matrigel matrix solution
Manufactured by BD
Sourced in United States
Matrigel matrix solution is a gelatinous protein mixture derived from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells. It is a complex extracellular matrix (ECM) that provides a basement membrane-like environment for cell growth and differentiation. The solution is used as a substrate for the in vitro culture of various cell types, including epithelial, endothelial, and stem cells.
Automatically generated - may contain errors
Lab products found in correlation
7 protocols using matrigel matrix solution
1
Cell Invasion Assay Using Transwell Inserts
2
Transwell Invasion and Migration Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The invasion ability was measured using a transwell chamber consisting of a 24-well plate with membrane inserts (Corning, New York, USA) containing polycarbonate filters (pore size: 8 μm) that were precoated with 60 µL of 1 mg/mL Matrigel matrix solution (BD Biosciences, California, USA). A suspension of 8 × 104 cells in 200 µL of serum-free culture medium was added to the inserts, which were then placed in the lower chamber containing 600 µL of 10 % FBS culture medium. The cells were left to migrate through the membranes for 24 h prior to fixing with 4 % paraformaldehyde and staining with crystal violet. Images were captured using the EVOS FL Color Imaging System (Thermo Fisher Scientific). The cell migration assay was performed in the same manner, except that the inserts were not coated with Matrigel.
3
Transwell Invasion Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The invasion assay was performed in a transwell chamber consisting of a 24‐well plate with membrane inserts (Corning, New York, USA) containing 8‐mm‐pore‐sized polycarbonate filters precoated with 60 μl of 1 mg/ml Matrigel matrix solution (BD Biosciences, San Jose, USA). A suspension of 4 × 104 cells in 200 μl of serum‐free culture medium was added to the inserts, and each insert was placed in the lower chamber containing 600 μl of 10% fetal bovine serum (FBS) culture medium. After 24 h, the cells that penetrated the membrane were fixed with 4% paraformaldehyde and stained with crystal violet. Images were captured by using the Evos Fl Color Imaging System (Thermo Fisher Scientific, Waltham, USA).
4
Cell Invasion Analysis via Transwell
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To conduct a cell invasion analysis, Matrigel matrix solution (BD Biosciences) diluted with 1640 was placed in 24‐pore Transwell Millipore and incubated for 37°C until gel was formed. 5 × 104 HTR‐8/SVneo cells were collected, diluted with 200 μL serum‐free medium, and inoculated in the superior ventricle. The lower chamber was added with 500 μL 15% FBS medium. After incubation at 37°C for 36 h, the chamber was taken out and washed twice with PBS. The cells were fixed in methanol for 20 min and stained with crystal violet. Under the light microscope, the cells were counted in five random fields with a magnification of 200 times.
5
Placental Villous Tissue Explant Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The placental villous tissues were dissected into explants of 2–5 mm in diameter and explanted as previously described (Chen et al., 2018 (link); Zhang et al., 2012 (link)). A 24‐well plate was precoated with 50 μl of a 1 mg/ml Matrigel matrix solution (CA# 354234, BD Biosciences, San Jose, CA, USA) and incubated at 37°C for 4 h for solidification. Serum‐free DMEM/F12 (Gibco) medium containing 10% FBS (PAN) with 20 mM NAM (#HY‐B0150, MedChemExpress, USA), 3 μM SRT1720 (#HY‐10532, MedChemExpress, USA), 500 nmol/l lentiviral vector‐based shRNA targeting SIRT1 or an equal concentration of scrambled shRNA (GeneChem, Shanghai, China) was added to the wells; the explants were cultured in 3% oxygen and 5% CO2 for 48 h. The explants with good attachment and outgrowth on the gel were assessed after 24 and 48 h, respectively. The evaluation of villus outgrowth was performed as previously described (Li et al., 2014 (link)).
6
Transwell Invasion Assay of HTR8/SVneo Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Invasion assays were conducted using 24-well transwell chambers (Falcon, USA) with 8 μm pore size and precoated with 60 μL of 1 mg/mL Matrigel matrix solution per chamber (BD Biosciences, USA). A suspension of 5×10 4 transfected HTR8/SVneo cells in 200 μL of serum-free culture medium was added to the upper transwell chambers, while the lower chambers were loaded with 600 μL of culture medium containing 10% FBS. After 24 h, the chambers were fixed with 4% paraformaldehyde and stained with crystal violet boric acid. Noninvading HTR8/SVneo cells were gently removed from the upper side of the membrane with a sterile cotton swab. Images were captured using the Evos FL Color Imaging System. The Evos FL Color Imaging System was used to capture images, and ImageJ 1.53o software was used to count invaded cells. Each experiment was performed in triplicate.
7
Matrigel-Based Cell Invasion Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The invasion assay was performed as previously reported by our laboratory (Yang et al., 2020 (link)). First, transwell inserts containing a polycarbonate membrane with an 8 μm pore size (Corning, New York, USA) were placed into 24‐well plates. The upper chamber was precoated with 60 μl of 1 mg/ml Matrigel matrix solution (BD Biosciences) before 8x104 cells in 200 μl of serum‐free culture medium were added. The lower chamber contained 600 μl of culture medium supplemented with 10% FBS. After incubation for 24 hr, the inserts were fixed with 4% paraformaldehyde and stained with crystal violet. Images were captured using the Evos Fl Color Imaging System (Thermo Fisher Scientific). All data were analyzed using Image J 1.50i software.
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!