For formation of trophoblast spheroids, 4 × 105 Sw.71-GFP cells were seeded into each well of a 96-well ultra-low attachment plate (Sigma-Aldrich) and centrifuged at 400 g for 5 minutes to cluster the cells together. The plate was incubated for 48 hours to allow the clumped trophoblast to form into a spheroid. The spheroids were individually collected and transferred onto other plates for migration or invasion assays. This in vitro model of trophoblast implantation is described in full by You et al.15 (link)
96 well ultra low attachment plate
Manufactured by Merck Group
The 96-well ultra-low attachment plate is a laboratory equipment designed for cell culture applications. It features a non-adherent surface that promotes the formation of cellular aggregates, such as spheroids and organoids, without the cells adhering to the plate.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
Defined trypsin inhibitor, by Thermo Fisher Scientific (1 mentions)
Tryple express, by Merck Group (1 mentions)
Rho kinase inhibitor y27632, by STEMCELL (1 mentions)
Laminin 521 coated culture dishes, by BioLamina (1 mentions)
Essential 8 medium, by Thermo Fisher Scientific (1 mentions)
Tryple express, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
Penicillin streptomycin, by STEMCELL (1 mentions)
2 protocols using 96 well ultra low attachment plate
1
Trophoblast Spheroid Formation Protocol
2
Culturing Fibroblasts and Induced Pluripotent Stem Cells
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Fibroblasts were cultured in DMEM (Sigma, cat no: D5796), 10% fetal bovine serum (ThermoFisher-Scientific, cat no: 10500056), 2 mM GlutaMAX™ (ThermoFisher-Scientific, cat no: 35050038), 1% penicillin/streptomycin (ThermoFisher-Scientific, cat no: 15140122) in a humidified atmosphere with 5% CO 2 at 37 • C and passaged using Try-pLE™ Express (ThermoFisher-Scientific, cat no: 12604039), and Defined Trypsin Inhibitor (ThermoFisher-Scientific, cat no: R007100). Human iPS cells were cultured in Essential-8™ medium (ThermoFisher Scientific, cat no: A1517001), on laminin-521 coated culture dishes (BioLamina; 5% CO2, 37 • C) and passaged as single cells using TrypLE™ Express and Defined Trypsin Inhibitor at a ratio of 1:10 when 80% confluence was reached.
For the embryoid body (EB) differentiation assay, iPSCs were dissociated with TrypLE™-Express, seeded into a 96-well ultra-low attachment plate (Sigma) in DMEM/F12, 20% Knock-out serum replacement, 3% FBS, 2 mM GlutaMAX™, 1x non-essential amino acids and 1% penicillin/streptomycin, supplemented with 10 μM Rho-kinase inhibitor Y27632 (Stem Cell Technologies). The next day, formed EBs were transferred to non-adherent culture plates for a total of 28 days of differentiation.
For the embryoid body (EB) differentiation assay, iPSCs were dissociated with TrypLE™-Express, seeded into a 96-well ultra-low attachment plate (Sigma) in DMEM/F12, 20% Knock-out serum replacement, 3% FBS, 2 mM GlutaMAX™, 1x non-essential amino acids and 1% penicillin/streptomycin, supplemented with 10 μM Rho-kinase inhibitor Y27632 (Stem Cell Technologies). The next day, formed EBs were transferred to non-adherent culture plates for a total of 28 days of differentiation.
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