Comprehensive Cell Surface Protein Profiling Identifies Specific Markers of Human Naive and Primed Pluripotent States.
Human pluripotent stem cells (PSCs) exist in naive and primed states and provide important models to investigate the earliest stages of human development. Naive cells can be obtained through primed-to-naive resetting, but there are no reliable methods to prospectively isolate unmodified naive cells during this process. Here we report comprehensive profiling of cell surface proteins by flow cytometry in naive and primed human PSCs. Several naive-specific, but not primed-specific, proteins were also expressed by pluripotent cells in the human preimplantation embryo. The upregulation of naive-specific cell surface proteins during primed-to-naive resetting enabled the isolation and characterization of live naive cells and intermediate cell populations. This analysis revealed distinct transcriptional and X chromosome inactivation changes associated with the early and late stages of naive cell formation. Thus, identification of state-specific proteins provides a robust set of molecular markers to define the human PSC state and allows new insights into the molecular events leading to naive cell resetting.
Epigenetics Programme, The Babraham Institute, Cambridge CB22 3AT, UK; Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, Cambridge CB2 1QR, UK.
Amanda J
AJ
Collier
Department of Clinical Science, Intervention, and Technology, Karolinska Institutet, 14186 Stockholm, Sweden; Division of Obstetrics and Gynecology, Karolinska Universitetssjukhuset, 14186 Stockholm, Sweden.
Sarita P
SP
Panula
Department of Clinical Science, Intervention, and Technology, Karolinska Institutet, 14186 Stockholm, Sweden; Division of Obstetrics and Gynecology, Karolinska Universitetssjukhuset, 14186 Stockholm, Sweden.
John Paul
JP
Schell
Nuclear Dynamics Programme, The Babraham Institute, Cambridge CB22 3AT, UK.
Peter
P
Chovanec
Department of Clinical Science, Intervention, and Technology, Karolinska Institutet, 14186 Stockholm, Sweden; Division of Obstetrics and Gynecology, Karolinska Universitetssjukhuset, 14186 Stockholm, Sweden.
Alvaro
A
Plaza Reyes
Department of Clinical Science, Intervention, and Technology, Karolinska Institutet, 14186 Stockholm, Sweden; Division of Obstetrics and Gynecology, Karolinska Universitetssjukhuset, 14186 Stockholm, Sweden.
Sophie
S
Petropoulos
Nuclear Dynamics Programme, The Babraham Institute, Cambridge CB22 3AT, UK.
Anne E
AE
Corcoran
Flow Cytometry Core Facility, The Babraham Institute, Cambridge CB22 3AT, UK.
Rachael
R
Walker
Center for Hematology and Regenerative Medicine, Department of Medicine, Karolinska Institute, 14186 Stockholm, Sweden.
Iyadh
I
Douagi
Department of Clinical Science, Intervention, and Technology, Karolinska Institutet, 14186 Stockholm, Sweden; Division of Obstetrics and Gynecology, Karolinska Universitetssjukhuset, 14186 Stockholm, Sweden. Electronic address: fredrik.lanner@ki.se.
Fredrik
F
Lanner
Epigenetics Programme, The Babraham Institute, Cambridge CB22 3AT, UK; Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, Cambridge CB2 1QR, UK; Centre for Trophoblast Research, University of Cambridge, Cambridge CB2 3EG, UK. Electronic address: peter.rugg-gunn@babraham.ac.uk.
Peter J
PJ
Rugg-Gunn
D000943:Antigens, Differentiation / Q000096:biosynthesis*; D002460:Cell Line; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D008565:Membrane Proteins / Q000096:biosynthesis*; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*
D000943:Antigens, Differentiation; D008565:Membrane Proteins
antibody library; blastocyst; cell surface markers; differentiation; embryonic stem cells; pluripotency; reprogramming