A fast and efficient size separation method for haploid embryonic stem cells
Open access
Datum
2017-09Typ
- Journal Article
Abstract
Hemizygous mutations introduced in haploid genomes can directly expose a phenotype, thus facilitating gene function analysis and forward genetic screening. Recently, mammalian haploid cells could be derived from mouse, rat, monkey, and human embryos and have been applied to screens of cellular mechanisms including cell signaling, pathogen host factors, and developmental pathways. Notably, haploid cell cultures have an intrinsic tendency for diploidization and, thus, require periodic cell sorting. Here, we report a method for rapid purification of haploid mouse embryonic stem cells from mixed cell populations with high viability and yield. Our method uses membranes with micrometer pores for force-free separation and facilitates enrichment of haploid cells without flow cytometry. The separation method simplifies maintaining haploid cell cultures and has further applications in establishing haploid cell lines from embryos and isolating cell cycle phases of mammalian cells. Mehr anzeigen
Persistenter Link
https://doi.org/10.3929/ethz-b-000211869Publikationsstatus
publishedExterne Links
Zeitschrift / Serie
BiomicrofluidicsBand
Seiten / Artikelnummer
Verlag
American Institute of PhysicsOrganisationseinheit
03978 - Wutz, Anton / Wutz, Anton
Förderung
152814 - X chromosome reactivation in development and transformation (SNF)
145026 - Forward genetic screening in mouse haploid embryonic stem cells (ESCs) to functionally dissect pathways in mammalian development and disease (SNF)