MMECs were seeded on 12-well plates (Greiner Bio-One) precoated with 50 μg/ml of Laminin-111 (BioLamina, LN111-02) at 4°C overnight, and incubated for 24h in MMEC growth media. MMECs were treated with 500μM 6-AN (Cayman Chemical Company, 10009315), or DMSO (Sigma-Aldrich, D2650) for 24h and used to assess cell viability, or trypsinised, counted and plated in mammosphere culture or used in fat pad transplantations. Live and Dead Cell Assay (Abcam, ab115347) was used for cell viability studies according to manufacturer’s instructions. Samples were imaged using a Nikon Eclipse Ts2 microscope with CFI Ph1 ADL 20× objective, NA 0.4, and CoolLED pE-300 white light source and DS-Qi1 camera (Nikon), and cells were counted using ImageJ 1.51 (NIH) in 2-3 images per group. For mammosphere culture, cells were plated in 1% methylcellulose (15 cP, Sigma-Aldrich, M7027) in MMEC growth media on 96-well low adhesion plates (Corning, 3474) at 1,000 cells per well. Mammospheres were counted 14 days after plating.
Laminin 111
Manufactured by BioLamina
Sourced in Sweden
Laminin-111 is a key structural component of the extracellular matrix. It serves as a substrate for cell attachment and provides a foundation for cell growth and differentiation.
Automatically generated - may contain errors
Lab products found in correlation
Live and dead cell assay, by Abcam (2 mentions)
Coolled pe 300, by Nikon (2 mentions)
Cfi ph1 adl 20 objective, by Nikon (2 mentions)
Ds qi1 camera, by Nikon (2 mentions)
M7027, by Corning (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
12 well plate, by Greiner (2 mentions)
Vitronectin, by Merck Group (1 mentions)
Matrigel matrix, by Corning (1 mentions)
4 protocols using laminin 111
1
Assessing MMEC Viability and Mammosphere Formation
2
Assessing MMEC Viability and Mammosphere Formation
3
Scaffold Preparation and Functionalization
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3D box-pore scaffolds were cut into 9 mm discs and placed into a 15.6 mm culture dish (24-well plate) (Waltham, Massachusetts, MA, USA) and 3D-aligned microfiber tracts were placed into a 34.8 mm culture dish (6-well plate) (Corning, New York, NY, USA) then sterilized with UV-light for 30 min, incubated with 1 M NaOH for 10 min and finally washed five times with 1x PBS. For functionalization, scaffolds or aligned microfiber tracts were incubated with 10 µg mL−1 laminin-111 (BioLamina, Sundbyberg, Sweden) at 4 °C overnight.
4
Cell Culture Substrate Enrichment Protocol
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Prior cell culture the mats were enriched for 3 h at 4 °C with either Matrigel Matrix (20 × dilution, Corning), laminin-111 (concentration 25 µg/ml, BioLamina) or vitronectin (concentration 12.5 µg/ml, Sigma-Aldrich) in hepatocyte culture medium. Control mats were incubated in culture medium alone. After the incubation the mats were briefly washed with phosphate buffered saline (PBS) pH 7.4 before seeding the cells.
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