title: Isolation and cultivation of naive-like human pluripotent stem cells based on HERVH expression.
abstract: The ability to derive and stably maintain ground-state human pluripotent stem cells (hPSCs) that resemble the cells seen in vivo in the inner cell mass has the potential to be an invaluable tool for researchers developing stem cell-based therapies. To date, derivation of human naive-like pluripotent stem cell lines has been limited to a small number of lineages, and their long-term culturing remains problematic. We describe a protocol for genetic and phenotypic tagging, selecting and maintaining naive-like hPSCs. We tag hPSCs by GFP, expressed by the long terminal repeat (LTR7) of HERVH endogenous retrovirus. This simple and efficient protocol has been reproduced with multiple hPSC lines, including embryonic and induced pluripotent stem cells, and it takes ∼6 weeks. By using the reporter, homogeneous hPSC cultures can be derived, characterized and maintained for the long term by repeated re-sorting and re-plating steps. The HERVH-expressing cells have a similar, but nonidentical, expression pattern to other naive-like cells, suggesting that alternative pluripotent states might exist.
authors.affiliation: Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.
authors.forename: Jichang
authors.initials: J
authors.lastname: Wang
authors.affiliation: Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.
authors.forename: Manvendra
authors.initials: M
authors.lastname: Singh
authors.affiliation: Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.
authors.forename: Chuanbo
authors.initials: C
authors.lastname: Sun
authors.affiliation: Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Besser
authors.affiliation: Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Prigione
authors.affiliation: Paul Ehrlich Institute, Division of Medical Biotechnology, Langen, Germany.
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Ivics
authors.affiliation: Department of Biology and Biochemistry, University of Bath, Bath, UK.
authors.forename: Laurence D
authors.initials: LD
authors.lastname: Hurst
authors.affiliation: Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin, Germany.
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Izsvák
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D020077:Endogenous Retroviruses / Q000235:genetics; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006801:Humans; D039904:Pluripotent Stem Cells / Q000502:physiology*; D013194:Staining and Labeling
chemical_list: D049452:Green Fluorescent Proteins