Cell invasion ability was observed by spheroid collagen type I invasion assay and tested by Transwell invasion assay based on published protocol [29, (link)30] (link). For spheroid collagen type I invasion assay, HUVECs at a total number of 5.0 × 10 4 in 100 μL EGM-2 medium (Lonza) were seeded on 96-well ultra-low attachment surface plates (Corning) for 2 days to generate cell spheres. Collagen type I (C8062, Solarbio) solution with a final concentration of 1 mg/mL was prepared according to the manufacturer's instructions. The spheres were then embedded in the collagen gel, and 100 μL complete ECM with or without additional VEGF at 50 ng/ mL was added. Invasion of spheres was observed the next day under a microscope (CKX41, Olympus) with a CCD camera (DP70, Olympus). For Transwell invasion assay, the upper chamber of Transwell chamber (Corning) was precoated with 100 μL Matrigel Basement Membrane Matrix (matrigel) (1:20, BD Biosciences) overnight at 37 °C and then cultivated with HUVECs at a total number of 1.0 × 10 5 in 200 μl ECM containing 0.5% FBS without ECGS. 500 μL complete ECM with 50 ng/ml VEGF-165 (Sino Biological) was added to the lower chamber. Cells were cultured for 12 h and were fixed by 4% PFA and stained with crystal violet. Number of cells invading to the lower chamber was observed under a microscope (CKX41; Olympus) with a CCD camera (DP70; Olympus).
C8062
Manufactured by Solarbio
Sourced in China
The C8062 is a laboratory equipment product. It is designed for a specific core function, but a detailed description while maintaining an unbiased and factual approach is not available.
Automatically generated - may contain errors
5 protocols using c8062
1
Evaluating Cell Invasion Potential
2
Collagen Gel-Based Cell Contraction Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After stimulation, cells were trypsinized, counted, and mixed with the Collagen Gel Working Solution prepared according to the manufacturer’s protocol (C8062, Solarbio). The solution was added to 24 well plates and allowed to polymerize for 20 min at 37 °C. Fresh growth medium was added to the solidified collagen gels, and plates were returned to the incubator. After OGD for 6 h, the surface of contraction was released and contraction was monitored. Next, the surface area of contracted gels was imaged using a Chemi Doc instrument (Bio-Rad) and measured using the ImageJ software (NIH).
3
Cell Adhesion Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In cell adhesion assay, 24 well plates were coated with the rat tail type I collagen (C8062, Solarbio, China), washed with PBS 2-3 times, and then placed at 37 ℃ overnight. The treated cell suspension (about 50000 cells) was planted on the coated culture plate and placed in the 37 ℃ cell incubator for 30 minutes and 1 hour respectively. The liquid was removed and washed with PBS for 1-2 times. First, the cells were xed with 4% paraformaldehyde solution for 20 minutes, washed gently for 1-2 times, and then added crystal violet staining solution, dyed for 20 minutes, gently washed 1-2 times, and nally taken photos in 3 areas randomly selected under the microscope.
4
Rat Tail Collagen Hydrogel Migration
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HVF cell-laden rat tail collagen (C8062, Solarbio) clots were prepared as previously described. 5 μL rat tail collagen solution (2 mg mL−1, 0.01MPBS) containing 1.5 × 105 cells was dropped on Teflon plate via a 1 mL syringe and placed in 37 °C incubator for 25 min. The clots were then immersed into different hydrogel precursors followed by photo-immobilization with 10 mW cm−2 365 nm lamp for 30 s. After 14 days, cytoskeleton staining was performed as mentioned above and both confocal microscopy and the light microscope were used to analyze 3D cell migration behaviors. The distance of migrating cells and the migration area was measured by Image J software.
5
Collagen Gel Contraction Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HConFs were harvested by exposure to trypsin-EDTA, washed twice, and resuspended in serum-free DMEM/F12 medium. Type I collagen from rat tail tendon (C8062, 5 mg/ml; Solarbio Science and Technology Co., Ltd., Beijing, China), 10× PBS, 0.06 × 0.1 mol/L NaOH, and HConFs suspension were mixed on ice in an appropriate volume ratio (final concentration of type I collagen, 1 mg/ml; final cell density, 2.5 × 105 cells/ml). A portion (0.5 ml) of the mixture was added to each well of the 24-well culture plates and allowed to solidify by incubation at room temperature for 20 min. The collagen gels were freed from the sides of the wells with a sterilized 10-μL pipette. Serum-free medium (0.5 ml) containing 10 ng/ml TGF-β1 or 5 µM AZD6738 was added to the surface of each corresponding gel. Represent images of collagen gel contraction were photographed by a Fluor ChemE (92–14860–00; ProteinSimple, San Jose, CA, United States). The gel size was measured by ImageJ. The percentage of contraction area of each group was calculated as follows: (initial gel area - gel area at each time point)/initial gel area × 100% (Lan et al., 2021 (link)).
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!