Matrigel 354230

Advanced Dulbecco’s modified Eagle’s medium/F12, Antibiotic-Antimycotic (Anti-Anti), and B-27 supplement were from Thermo Fisher Scientific (Waltham, MA); recombinant human R-spondin-1, recombinant human EGF, recombinant human HGF, recombinant human FGF10, and recombinant human noggin were from PeproTech (Cranbury, NJ):, gastrin, N-acetylcysteine, Y27632, nicotinamide and A83-01 were from Millipore-Sigma (Burlington, MA); human recombinant Wnt3A, Forskolin and human recombinant IL-17A were from R&D Systems (Minneapolis, MN). Matrigel (354230) and cell recovery solution were from Corning (Kennebunk, ME, USA).

The wells of a 96-well plate were coated with 50 μl of nothing (control group), MeSF hydrogel, and Matrigel 354230 (Corning^®, a basement membrane matrix derived from the Engelbreth-Holm-Swarm mouse sarcoma used as a positive control). The Matrigel 354230 solidified when incubated at 37°C for 30 minutes. Then, 100 μl of the HUVECs suspension (4 × 10⁵ cells/mL) were added to each well of the 96-well plate. The plate was incubated at 37°C in a humidified incubator with 5% CO₂ for 72 hours. The cells were imaged using an inverted microscope at set time intervals. The formation of tube-like structures by the cells was analyzed using Image J 1.52q1.52v.