Primary/secondary spheroid enrichment analysis was done to compare the frequencies of primary spheroids formed from short-term cell tumor cultures compared to secondary spheroids formed from dissociated primary spheroids in order to assess the self-renewal capacity of the enriched CSC. Equal numbers of cells derived from short-term HNSCC CSC cultures (primary) and cells derived from primary enriched spheroids (secondary) were suspended in SC medium on ultra-low attachment six-well plates (Corning, New York). To form secondary spheroids, primary non-adherent spheroids were selected using a 37 μm Reversible Strainer (STEMCELL Technologies Inc., Vancouver, British Columbia, Canada), centrifuged at 800×g for 4 min, and then mechanically dissociated and re-plated. Large spheroids (more than 100 cells) from primary and secondary cultures were counted and photographed using a phase contrast photomicroscope (Leica Microsystems, Wetzlar, Germany).
37 μm reversible strainer
Manufactured by STEMCELL
The 37 μm Reversible Strainer is a laboratory instrument designed for size-based filtration. It features a 37 μm mesh that can be used to separate particles or cells based on their size. The strainer can be easily reversed for backflushing or cleaning purposes.
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Lab products found in correlation
Phase contrast photomicroscope, by Leica (1 mentions)
Ultra low attachment six well plate, by Corning (1 mentions)
Stemdiff neural induction medium smadi, by STEMCELL (1 mentions)
Stemdiff neural rosette selection reagent, by STEMCELL (1 mentions)
Stemdiff smadi neural induction kit, by STEMCELL (1 mentions)
Accutase, by Merck Group (1 mentions)
Stemdiff neural progenitor medium, by STEMCELL (1 mentions)
Mtesr1 media, by STEMCELL (1 mentions)
2 protocols using 37 μm reversible strainer
1
Comparative Analysis of Primary and Secondary Spheroid Formation
2
Efficient Directed Differentiation of PSCs into NPCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Primed PSCs were differentiated into expandable NPCs by using the STEMdiff SMADi Neural Induction Kit (Stem Cell Technologies) as previously described [34 (link)–36 (link)]. In brief, primed PSCs were maintained on a Matrigel (Corning)-coated plate in mTeSR1 media (Stem Cell Technologies) prior to the NPC induction. The cells were harvested using Accutase (EMD Millipore) and transferred at 3 x 106 cells to a well of an AgrreWell800 plate (Stem Cell Technologies) in STEMdiff Neural Induction Medium + SMADi (Stem Cell Technologies) supplemented with 10 μM Y-27632. Five days later, uniformly sized aggregates were collected using a 37 μm Reversible Strainer (Stem Cell Technologies) and plated onto a Matrigel-coated 6-well plate in STEMdiff Neural Induction Medium + SMADi. Seven days later, neural rosette structures were selectively removed by using STEMdiff Neural Rosette Selection Reagent (Stem Cell Technologies) and plated onto a new Matrigel-coated 6-well plate in STEMdiff Neural Induction Medium + SMADi. After that, the cells were passaged every 2–3 days until day 30 post-differentiation. The established NPCs were maintained on a Matrigel-coated plate in STEMdiff Neural Progenitor Medium (Stem Cell Technologies) and passaged every 3–4 days.
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