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Title:           The pluripotency state of human embryonic stem cells derived from single blastomeres of eight-cell embryos
Subject:         Cells & Development, 179 (2024) 203935. doi:10.1016/j.cdev.2024.203935
Keywords:        hESC,Derivation,Blastomeres,Naïve,Primed,Pluripotency
Author:          Ot Massafret
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      Title: The pluripotency state of human embryonic stem cells derived from single blastomeres of eight-cell embryos
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        Title: 1 Introduction
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        Title: 2 Materials and methods
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         Item: 
          Title: 2.1 Embryo thawing and culture
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          Title: 2.2 Feeder cells culture and inactivation
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          Title: 2.3 Derivation and culture of hESCs
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          Title: 2.4 Conversion of a pre-existing primed hESC line
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          Title: 2.5 Characterization of hESC lines
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            Title: 2.5.1 Immunostaining for intracellular markers
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            Title: 2.5.2 Immunostaining for cell surface marker SSEA3
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            Title: 2.5.3 Karyotyping
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            Title: 2.5.4 Alkaline phosphatase assay
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          Title: 2.6 Clonogenicity and doubling time assays
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          Title: 2.7 TFE3 intracellular localization analysis
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          Title: 2.8 Mitochondrial activity analysis
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          Title: 2.9 DNA extraction and 5mC/5hmC quantification
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          Title: 2.10 Embryoid body formation
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          Title: 2.11 RNA extraction and real-time quantitative PCR
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          Title: 2.12 Library preparation and RNA-sequencing
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          Title: 2.13 RNA-seq processing and analysis
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          Title: 2.14 Experimental design
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          Title: 2.15 Statistical analysis
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        Title: 3 Results
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          Title: 3.1 Generation of outgrowths and PICMIs from single blastomeres and whole blastocysts
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          Title: 3.2 Effects of GSK3βi, ROCKi and bFGF on hESC derivation efficiencies from single blastomeres and whole blastocysts
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          Title: 3.3 Characterization of hESC lines
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          Title: 3.4 hESC morphology and behaviour in culture
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          Title: 3.5 Analysis of naïve pluripotency indicators
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          Title: 3.6 Expression of naïve and primed pluripotency genes
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          Title: 3.7 Transcriptome analysis
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          Title: 3.8 Differential expression gene analysis
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          Title: 3.9 Primary germ layer differentiation capacity of hESCs by EBs formation
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          Title: 3.10 hESC derivation from single blastomeres in naïve conditions
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        Title: 4 Discussion
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        Title: 5 Conclusion
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       Item: 
        Title: Funding
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       Item: 
        Title: CRediT authorship contribution statement
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       Item: 
        Title: Declaration of competing interest
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        Title: Data availability
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        Title: Acknowledgements
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        Title: Appendix A Supplementary data
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        Title: References
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       Flags: Print
      Annotation: 
       Subtype: Link
       Rect: 241, 668, 286, 674
       Flags: Print
      Annotation: 
       Subtype: Link
       Rect: 49, 660, 142, 666
       Flags: Print
      Annotation: 
       Subtype: Link
       Rect: 78, 636, 210, 643
       Flags: Print
      Annotation: 
       Subtype: Link
       Rect: 462, 732, 539, 738
       Flags: Print
      Annotation: 
       Subtype: Link
       Rect: 318, 724, 372, 730
       Flags: Print
      Annotation: 
       Subtype: Link
       Rect: 318, 692, 446, 698
       Flags: Print
      Annotation: 
       Subtype: Link
       Rect: 354, 660, 468, 666
       Flags: Print
      Annotation: 
       Subtype: Link
       Rect: 431, 644, 507, 651
       Flags: Print
      Annotation: 
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       Rect: 318, 636, 380, 643
       Flags: Print
  Checksum: ccb5c139
   Type: CRC32
  Checksum: 2fae643c436b053b5f454354d68f4a3e
   Type: MD5
  Checksum: 9ba11cab5159160642e47c5bc645c164a242cb4b
   Type: SHA-1
  Checksum: 5e51e86a8466808cb4354d4ca32c30a5200217b8c176be5fce9874640509aaaa
   Type: SHA-256