The biomimetic microfluidic microvessel analogue was fabricated using PDMS lithography. Briefly, the basic and curing agent of PDMS was mixed at 10:1 ratio and poured onto patterned mold wafer. Then, the patterned PDMS layer was irreversibly bonded onto a glass slide by plasma treatment. The assembled microfluidic device was placed in the 65°C overnight to promote the bonding between PDMS and glass slide. Type I collagen gel (Corning Inc.) isolated from rat tail was introduced into the central compartment as the ECM component and polymerized at 37˚C in a humidified incubator overnight prior to cell seeding. High molecular weight HA was mixed with collagen gel at a concentration of 1 mg/mL HA to prepare collagen/HA matrices. The collagen-only or the collagen/HA pre-polymer mixture was pipetted into the central channel and polymerized overnight in an incubator before further usage.
Type 1 collagen gel
Manufactured by Corning
Type I collagen gel is a laboratory product derived from natural collagen. It serves as a three-dimensional extracellular matrix scaffold for cell culture applications.
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Lab products found in correlation
4 protocols using type 1 collagen gel
1
Biomimetic Microvessel Fabrication
2
3D Cell Invasion Assay Protocol
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For 3D cell invasion assays, 105 cells (BT549 or 4T1)
were embedded in 20 μl of type I collagen gel (2.2 mg/ml, Corning).
After gelling, the plug was embedded in a cell-free collagen gel (2.2 mg/ml)
within a 24-well plate. After allowing the surrounding collagen matrix to
gel (1 h at 37°C), 0.5 ml of culture medium was added to the top of
the gel and cultured for another two days. Invasion distance from the inner
collagen plug into the outer collagen gel was quantified.
were embedded in 20 μl of type I collagen gel (2.2 mg/ml, Corning).
After gelling, the plug was embedded in a cell-free collagen gel (2.2 mg/ml)
within a 24-well plate. After allowing the surrounding collagen matrix to
gel (1 h at 37°C), 0.5 ml of culture medium was added to the top of
the gel and cultured for another two days. Invasion distance from the inner
collagen plug into the outer collagen gel was quantified.
3
3D Cell Invasion Assay Protocol
4
Hepatobiliary Organoid Induction Protocol
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A step-by-step protocol describing induction of the HBTO can be found at Protocol Exchange32 . Cholangiocytes were resuspended in the growth medium and plated in 24-well plates coated with 200 μl of 4 mg/ml type I collagen gel prepared from high concentration type I collagen (Corning) at a density of 50,000 cells/well. Five days after plating, SHs were added to each well at a density of 50,000 cells/well. Following a further two days of incubation, the medium was replaced with differentiation medium supplemented with 10 ng/ml OSM and then overlaid with collagen gel containing 20% MG (Col-MG), which was prepared by mixing 2 mg/ml type I collagen gel and MG (v/v = 4:1) on ice. The plate was incubated at 37 °C for 3–4 h to form a gel, and then the differentiation medium was added. Culture medium was replaced with fresh medium every four days.
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