Embryoid bodies (EBs) are three-dimensional aggregates comprised of human pluripotent stem cells (hPSCs). Within EBs, hPSCs undergo differentiation and cell specification along the three germ layers, which are commonly used as an assessment of the initial hPSCs' pluripotency. The protocol for formation of spin EBs can be used as a basis for the development of protocols for directed differentiation via EBs.
Confirmation of pluripotency by spin embryoid body formation
- 24-well plates, cell-culture treated, flat bottom
- 96-well plates, non-treated, V bottom
- Advanced RPMI 1640
(with glucose, sodium pyruvate, and non-essential amino acids; without L-glutamine and HEPES)
- B-27 Supplement (50X), serum free
- DEF-CS COAT-1
- GlutaMAX-I (100X; 200 mM)
- PBS Dulbecco's with Ca2+ & Mg2+ (D-PBS +/+)
- PBS Dulbecco's w/o Ca2+ & Mg2+ (D-PBS –/–)
(PEST; 10,000 units/ml of penicillin and 10,000 µg/ml of streptomycin)
- TrypLE Select Enzyme (1X), no phenol red
(Thermo Fisher Scientific, Cat. # 12563011)
User-generated protocols are based on internal proof-of-concept experiments, customer collaborations, and published literature. In some cases, relevant results are discussed in our research news BioView blog articles. While we expect these protocols to be successful in your hands, they may not be fully reviewed or optimized. We encourage you to contact us or refer to the published literature for more information about these user-generated and -reported protocols.
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