Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2023 Nov 14;12:RP87081. doi: 10.7554/eLife.87081

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2023, Mavrommatis et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

PMC Copyright notice

Figure 2. — (A) Stepwise brightfield images depicting delamination/migration of progenitor population during organoid culture progression and corresponding myofiber formation. (B) Gene ontology enrichment analysis comparing 4 and 8 wk organoids attributes muscle identity at 8 wk post differentiation and highlights muscle system development and neural lineage arrest among the top upregulated and downregulated gene ontology terms, respectively. (C–E) Organoid overview on day 35 indicates predominant expression of FastMyHC⁺ and PAX7⁺ myogenic populations, while SOX2⁺ neural populations demarcate SOX2 neural plate border epithelium location as observed at earlier stages (day 16) (C); PAX7 cells are of myogenic origin (PAX7⁺/SOX10^-), MF20⁺ myotubes are in their proximity and MYOD1⁺ cells appear at organoid periphery (D); TUJ1⁺ neurons are restricted to inner organoid areas and close to SOX2⁺ epithelium, while FastMyHC⁺ myofibers occupy exterior organoid areas (E). (F) Histographs based on FACS intracellular quantification depicting percentage of PAX7⁺ or MYOD1⁺ cells through differentiation protocol. For each replicate, 10 organoids were pooled, n = 10. Statistics: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns: not significant. (G) Heatmap of HOX gene cluster emphasizes organoid culture limb axial anatomical identity by depicting transition from an initial limb bud (HOX 9–10) toward a more distal identity (HOX 11–13) at 8 and 16 wk post differentiation, respectively. Scale bars, 500 μm (C), 200 μm (A, D, E).

Figure 2—figure supplement 1. — (A) Stepwise brightfield images depicting delamination/migration of progenitor population during organoid culture progression and corresponding myofiber formation. (B) Gene ontology enrichment analysis comparing 4 and 8 wk organoids attributes muscle identity at 8 wk post differentiation and highlights muscle system development and neural lineage arrest among the top upregulated and downregulated gene ontology terms, respectively. (C–E) Organoid overview on day 35 indicates predominant expression of FastMyHC⁺ and PAX7⁺ myogenic populations, while SOX2⁺ neural populations demarcate SOX2 neural plate border epithelium location as observed at earlier stages (day 16) (C); PAX7 cells are of myogenic origin (PAX7⁺/SOX10^-), MF20⁺ myotubes are in their proximity and MYOD1⁺ cells appear at organoid periphery (D); TUJ1⁺ neurons are restricted to inner organoid areas and close to SOX2⁺ epithelium, while FastMyHC⁺ myofibers occupy exterior organoid areas (E). (F) Histographs based on FACS intracellular quantification depicting percentage of PAX7⁺ or MYOD1⁺ cells through differentiation protocol. For each replicate, 10 organoids were pooled, n = 10. Statistics: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, ns: not significant. (G) Heatmap of HOX gene cluster emphasizes organoid culture limb axial anatomical identity by depicting transition from an initial limb bud (HOX 9–10) toward a more distal identity (HOX 11–13) at 8 and 16 wk post differentiation, respectively. Scale bars, 500 μm (C), 200 μm (A, D, E).