This protocol describes an EDTA-based passaging procedure to be used with chemically defined E8 medium that serves as a tool for basic and translational research into human pluripotent stem cells (iPSCs). In this protocol, passaging one six-well or 10 cm plate of cells takes about 6–7 min. This enzyme-free protocol achieves maximum cell survival without enzyme neutralization, centrifugation, or drug treatment. It also allows for higher throughput, requires minimal material and limits contamination. Here we describe how to produce a consistent E8 medium for routine maintenance and reprogramming and how to incorporate the EDTA-based passaging procedure into human induced PSC (iPSC) derivation, colony expansion, cryopreservation and teratoma formation. This protocol has been successful in routine cell expansion, and efficient for expanding large-volume cultures or a large number of cells with preferential dissociation of PSCs. Effective for all culture stages, this procedure provides a consistent and universal approach to passaging human pluripotent stem cells in E8 medium.
Rates of live birth or ongoing pregnancy after euploid SET (n¼987 transfers) did not differ by platform (Table ; p¼0.83).CONCLUSIONS: Euploid call rates differ as a function of PGT-A technology platform after adjusting for age and eggs collected. However, these differences do not seem to translate into different pregnancy outcomes after SET of a ''euploid'' blastocyst. Further investigation should attempt to reconcile these differences and clarify if and how advances in PGT-A technology platforms translate to patients.
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