TY - JOUR
T1 - Capture of Mouse and Human Stem Cells with Features of Formative Pluripotency
AU - Kinoshita, Masaki
AU - Barber, Michael
AU - Mansfield, William
AU - Cui, Yingzhi
AU - Spindlow, Daniel
AU - Stirparo, Giuliano Giuseppe
AU - Dietmann, Sabine
AU - Nichols, Jennifer
AU - Smith, Austin
N1 - Funding Information:
We thank Vicki Metzis for the ATAC-seq protocol, Elsa Sousa for help with Xist FISH, and Meng Amy Li for the gRNA expression vector. Tüzer Kalkan, Nicola Reynolds, and Laurence Bates advised on ChIP-seq. We are grateful to Maike Paramor, Vicki Murray, Peter Humphreys, Darran Clements, Andrew Riddell, Charles-Etienne Dumeau, and biofacility staff for technical support and to the CSCI core bioinformatics team for data processing and routine analysis. Sequencing was performed by the CRUK Cambridge Institute Genomics Core Facility. Shahzaib Ahmed and Benjamin Porteous contributed to experiments. Rosalind Drummond and James Clarke provided laboratory assistance. We thank Brian Hendrich for comments on the manuscript. The Cambridge Stem Cell Institute receives core funding from Wellcome and the Medical Research Council. This research was funded by the Biotechnology and Biological Sciences Research Council and the Medical Research Council United Kingdom. A.S. is a Medical Research Council professor. Conceptualization, A.S.; Methods, M.K.; Formal Analysis, M.B. D.S. G.G.S. and S.D.; Investigation, M.K. W.M. Y.C. and J.N.; Writing, M.K. and A.S.; Supervision, A.S. The authors declare no competing interests
Funding Information:
We thank Vicki Metzis for the ATAC-seq protocol, Elsa Sousa for help with Xist FISH, and Meng Amy Li for the gRNA expression vector. Tüzer Kalkan, Nicola Reynolds, and Laurence Bates advised on ChIP-seq. We are grateful to Maike Paramor, Vicki Murray, Peter Humphreys, Darran Clements, Andrew Riddell, Charles-Etienne Dumeau, and biofacility staff for technical support and to the CSCI core bioinformatics team for data processing and routine analysis. Sequencing was performed by the CRUK Cambridge Institute Genomics Core Facility. Shahzaib Ahmed and Benjamin Porteous contributed to experiments. Rosalind Drummond and James Clarke provided laboratory assistance. We thank Brian Hendrich for comments on the manuscript. The Cambridge Stem Cell Institute receives core funding from Wellcome and the Medical Research Council . This research was funded by the Biotechnology and Biological Sciences Research Council and the Medical Research Council United Kingdom . A.S. is a Medical Research Council professor.
Publisher Copyright:
© 2020 The Authors
PY - 2021/3/4
Y1 - 2021/3/4
N2 - Pluripotent cells emerge as a naive founder population in the blastocyst, acquire capacity for germline and soma formation, and then undergo lineage priming. Mouse embryonic stem cells (ESCs) and epiblast-derived stem cells (EpiSCs) represent the initial naive and final primed phases of pluripotency, respectively. Here, we investigate the intermediate formative stage. Using minimal exposure to specification cues, we derive stem cells from formative mouse epiblast. Unlike ESCs or EpiSCs, formative stem (FS) cells respond directly to germ cell induction. They colonize somatic tissues and germline in chimeras. Whole-transcriptome analyses show similarity to pre-gastrulation formative epiblast. Signal responsiveness and chromatin accessibility features reflect lineage capacitation. Furthermore, FS cells show distinct transcription factor dependencies, relying critically on Otx2. Finally, FS cell culture conditions applied to human naive cells or embryos support expansion of similar stem cells, consistent with a conserved staging post on the trajectory of mammalian pluripotency. Three stages of pluripotency have been proposed: naive, formative, and primed. Kinoshita and colleagues derived stem cells with properties anticipated for formative pluripotency by culturing mouse epiblast under conditions of low growth factor stimulation. Application to human embryos resulted in propagation of similar stem cells.
AB - Pluripotent cells emerge as a naive founder population in the blastocyst, acquire capacity for germline and soma formation, and then undergo lineage priming. Mouse embryonic stem cells (ESCs) and epiblast-derived stem cells (EpiSCs) represent the initial naive and final primed phases of pluripotency, respectively. Here, we investigate the intermediate formative stage. Using minimal exposure to specification cues, we derive stem cells from formative mouse epiblast. Unlike ESCs or EpiSCs, formative stem (FS) cells respond directly to germ cell induction. They colonize somatic tissues and germline in chimeras. Whole-transcriptome analyses show similarity to pre-gastrulation formative epiblast. Signal responsiveness and chromatin accessibility features reflect lineage capacitation. Furthermore, FS cells show distinct transcription factor dependencies, relying critically on Otx2. Finally, FS cell culture conditions applied to human naive cells or embryos support expansion of similar stem cells, consistent with a conserved staging post on the trajectory of mammalian pluripotency. Three stages of pluripotency have been proposed: naive, formative, and primed. Kinoshita and colleagues derived stem cells with properties anticipated for formative pluripotency by culturing mouse epiblast under conditions of low growth factor stimulation. Application to human embryos resulted in propagation of similar stem cells.
KW - chimaera
KW - epiblast
KW - formative pluripotency
KW - lineage induction
KW - pluripotent stem cell
KW - primordial germ cell
KW - self-renewal
UR - http://www.scopus.com/inward/record.url?scp=85097884694&partnerID=8YFLogxK
U2 - 10.1016/j.stem.2020.11.005
DO - 10.1016/j.stem.2020.11.005
M3 - Article
C2 - 33271069
AN - SCOPUS:85097884694
SN - 1934-5909
VL - 28
SP - 453-471.e8
JO - Cell Stem Cell
JF - Cell Stem Cell
IS - 3
ER -