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Series GSE252114 Query DataSets for GSE252114
Status Public on Jan 05, 2024
Title Self-renewing human naïve pluripotent stem cells dedifferentiate in 3D culture and form blastoids spontaneously
Organism Homo sapiens
Experiment type Expression profiling by high throughput sequencing
Summary Human naïve pluripotent stem cells (hnPSCs) can generate integrated models of blastocysts termed blastoids upon switch to inductive medium. However, the underlying mechanisms remain obscure. Here we report that self-renewing hnPSCs spontaneously and efficiently give rise to blastoids upon three dimensional (3D) suspension culture. The spontaneous blastoids mimic early stage human blastocysts in terms of structure, size, and transcriptome characteristics and are capable of progressing to post-implantation stages. This property is conferred by the glycogen synthase kinase-3 (GSK3) signalling inhibitor IM-12 present in 5iLAF self-renewing medium. IM-12 upregulates oxidative phosphorylation-associated genes that underly the capacity of hnPSCs to generate blastoids spontaneously. Starting from day one of self-organization, hnPSCs at the boundary of all 3D aggregates dedifferentiate into E5 embryo-like intermediates. Intermediates co-express SOX2/OCT4 and GATA6 and by day 3 specify trophoblast fate, which coincides with cavity and blastoid formation. In summary, spontaneous blastoid formation results from 3D culture triggering dedifferentiation of hnPSCs into earlier embryo-like intermediates which are then competent to segregate blastocyst fates.
 
Overall design We convert human primed pluripotent stem cells to a naïve state by GSK3 inhibitor CHIR99021 to IM-12 switch (CI-hnPSCs). We find that CI-hnPSCs spontaneously and efficiently generate blastocyst-like structures (blastoids) in the 5iLAF self-renewing medium. Removing BRAF and MEK inhibitors from 5iLAF, using 3iLAF from day 3 of blastoid formation results in hypoblast specification.
 
Contributor(s) Guo M, Wu J, Chen C, Wang X, Gong A, Guan W, Karvas RM, Wang K, Min M, Wang Y, Theunissen TW, Gao S, Silva JC
Citation(s) 38253551
Submission date Dec 27, 2023
Last update date Feb 09, 2024
Contact name Jinyi Wu
Organization name Guangzhou Laboratory
Street address No. 9 XingDaoHuanBei Road, Guangzhou International Bio Island
City Guangzhou
State/province Guangdong
ZIP/Postal code 510005
Country China
 
Platforms (2)
GPL24676 Illumina NovaSeq 6000 (Homo sapiens)
GPL30209 MGISEQ-2000RS (Homo sapiens)
Samples (164)
GSM8002709 RNA-seq of TJ 1# C-hnESC replicate 1
GSM8002710 RNA-seq of TJ 1# C-hnESC replicate 2
GSM8002711 RNA-seq of TJ 1# CI-hnESC replicate 1
Relations
BioProject PRJNA1061131

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE252114_RAW.tar 382.4 Mb (http)(custom) TAR (of MTX, TSV)
GSE252114_SpontaneousBlastoid_D0andD2_count.csv.gz 2.2 Mb (ftp)(http) CSV
GSE252114_TJ1_nESC_count.csv.gz 405.3 Kb (ftp)(http) CSV
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Raw data are available in SRA

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