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Agarose microgel culture delineates lumenogenesis in naive and primed human pluripotent stem cells
- Source :
- Stem Cell Reports
- Publication Year :
- 2021
- Publisher :
- Elsevier, 2021.
-
Abstract
- Summary Human periimplantation development requires the transformation of the naive pluripotent epiblast into a polarized epithelium. Lumenogenesis plays a critical role in this process, as the epiblast undergoes rosette formation and lumen expansion to form the amniotic cavity. Here, we present a high-throughput in vitro model for epiblast morphogenesis. We established a microfluidic workflow to encapsulate human pluripotent stem cells (hPSCs) into monodisperse agarose microgels. Strikingly, hPSCs self-organized into polarized epiblast spheroids that could be maintained in self-renewing and differentiating conditions. Encapsulated primed hPSCs required Rho-associated kinase inhibition, in contrast to naive hPSCs. We applied microgel suspension culture to examine the lumen-forming capacity of hPSCs and reveal an increase in lumenogenesis during the naive-to-primed transition. Finally, we demonstrate the feasibility of co-encapsulating cell types across different lineages and species. Our work provides a foundation for stem cell-based embryo models to interrogate the critical components of human epiblast self-organization and morphogenesis.<br />Graphical abstract<br />Highlights • High-throughput platform to encapsulate human PSCs in agarose microgels • Human PSCs self-organize into epiblast spheroids and undergo lumen expansion • Lumen expansion increases in primed versus naive human PSCs • Monodisperse co-culture of embryonic and extraembryonic stem cells<br />In this article, Boroviak and colleagues present a high-throughput platform to encapsulate human PSCs into agarose microgels. Human PSCs undergo lumenogenesis and self-organize into rosette-like structures, similar to the human peri-implantation embryo. Their microgel suspension culture regime provides a foundation for the controlled assembly of stem cell-based embryo models in our own species.
- Subjects :
- 0301 basic medicine
Resource
Cell type
Cell Survival
Induced Pluripotent Stem Cells
Morphogenesis
Cell Culture Techniques
Biology
Biochemistry
lumenogenesis
03 medical and health sciences
0302 clinical medicine
Spheroids, Cellular
Genetics
medicine
Humans
Cell Lineage
primed pluripotency
Induced pluripotent stem cell
microfluidic platform
Protein Kinase Inhibitors
rho-Associated Kinases
Microgels
epiblast
Sepharose
naive pluripotency
Spheroid
Embryo
Cell Differentiation
Cell Biology
Cells, Immobilized
self-organization
Epithelium
3. Good health
Cell biology
microgel suspension culture
030104 developmental biology
medicine.anatomical_structure
human development
Epiblast
embryonic structures
Stem cell
030217 neurology & neurosurgery
Germ Layers
Developmental Biology
epiblast spheroid
Subjects
Details
- Language :
- English
- ISSN :
- 22136711
- Volume :
- 16
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- Stem Cell Reports
- Accession number :
- edsair.doi.dedup.....147f0d63165d48a83852190a79672ddb