hiPSCs were plated at a concentration of 3.0 × 106 cells/well with E6 media into Aggrewell plates pretreated with 500 µl of Aggrewell Rinsing Solution (StemCell Technologies, Vancouver, BC). On day 4, embryoid bodies were replated into 35 mm Matrigel-coated immunofluorescence plates, and neural rosettes were fixed for imaging on day 8.
Aggrewell rinsing solution
Manufactured by STEMCELL
Sourced in France
Aggrewell Rinsing Solution is a specialized liquid used to rinse the Aggrewell culture plates. It is designed to facilitate the maintenance and preparation of these culture plates for cell culture applications.
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Lab products found in correlation
4 protocols using aggrewell rinsing solution
1
Differentiation of hiPSCs into Neural Rosettes
2
Generating Pluripotent Cell Spheroids
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Single cell suspensions of TRA-1-60 positive cells were deposited into AggreWell plates (Stem Cell Technologies) and allowed to aggregate into spheroids. Briefly, 1x106 cells positive for TRA-1-60 were suspended in 5 ml of EB Aggrewell medium (Stem Cell technologies #05893) and added into a well of an AggreWell400Ex plate (Stem Cell Technologies, # 27840) containing approximately 4,700 microwells with a diameter of 400 μm and pretreated with AggreWell rinsing solution (Stem Cell Technologies, # 07010). AggreWell plates were centrifuged at 100 x g for 3 minutes to deposit the cells in the microwells and incubated at 37°C for 24 hours, during which time spherical aggregates of deposited pluripotent cells (spheroids) were formed.
3
HepG2/C3A Spheroid Formation Protocol
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HepG2/C3A cell spheroids were prepared using an AggreWell™400 plate (Stemcell Technologies, Grenoble, France; Cat. no. 27845) according to manufactures specifications and as previously described (Fey & Wrzesinski, 2012a; Razian et al., 2013) . Wells were rinsed twice with 1 ml Aggrewell™ Rinsing solution (Stemcell Technologies, Grenoble, France; Cat. no. 07010). Air bubbles were removed from the well surface through centrifugation for 3 min at 3,000 xg. HepG2/C3A cells were seeded into each of the wells of the AggreWell™400 plate at a seeding density of 1.2 x 10 3 cells/well and centrifuged for 3 min at 120 xg. Following seeding, the cells were left in the AggreWell™ plate overnight to ensure aggregation and spheroid formation.
A c c e p t e d M a n u s c r i p t
A c c e p t e d M a n u s c r i p t
4
HepG2/C3A Spheroids Production
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HepG2/C3A cell spheroids were prepared using an AggreWell™400 plate (Stemcell Technologies, Grenoble, France) according to the manufacturers specifications and as previously published (Wrzesinski & Fey, 2012; Aucamp et al., 2017) . Wells were rinsed twice with 1 ml Aggrewell™
Rinsing solution (Stemcell Technologies, Grenoble, France), and HepG2/C3A cells were seeded into each of the wells of the AggreWell™ plate at a seeding density of 1.2 x 10 4 . Following centrifugation for 3 min at 120 x g, the plate was incubated overnight to allow spheroid formation.
Rinsing solution (Stemcell Technologies, Grenoble, France), and HepG2/C3A cells were seeded into each of the wells of the AggreWell™ plate at a seeding density of 1.2 x 10 4 . Following centrifugation for 3 min at 120 x g, the plate was incubated overnight to allow spheroid formation.
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