After embedding in Matrigel for 7–10 days, porcine enteroids were collected and incubated in 5 mM EDTA in PBS for 1 h at 4°C on a rotating platform. After this step, the samples were centrifuged at 400 g for 5 min and the supernatant was removed. The pellet was resuspended in enteroid growth medium on ultra-low attachment 24-well tissue culture plates (Corning); 10 µM Y27632 were also included in the media for the cultivation. The EDTA was used to disrupt the polymerization of the extracellular matrix (ECM) protein laminin [15 (link)]. ECM is a main component of Matrigel and it has been reported that ECM proteins can regulate the polarity of epithelial cells [39 (link)].
Ultra low attachment 24 well tissue culture plates
Manufactured by Corning
Ultra-low attachment 24-well tissue culture plates are designed to promote the formation of 3D cell cultures and spheroids. These plates have a proprietary surface treatment that minimizes cell attachment, encouraging the cells to aggregate and form self-organizing structures.
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Lab products found in correlation
2 protocols using ultra low attachment 24 well tissue culture plates
1
Isolating Porcine Enteroids from Matrigel
2
Anoikis Assay for Melanoma Cells
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To evaluate anoikis, melanoma cells expressing ASAH1 shRNA were seeded at 5 × 104 cells per well in ultra-low-attachment 24-well tissue culture plates (Corning) in DMEM or RPMI as appropriate for the cell line and incubated for 7 days at 37 °C. Live cells identified based on trypan blue dye exclusion were counted using a hemocytometer. Experiments were performed in triplicate.
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