Extended culture of 2D gastruloids to model human mesoderm development

Chen, Bohan; Khan, Hina; Yu, Zhiyuan; Yao, LiAng; Freeburne, Emily; Jo, Kyoung; Johnson, Craig; Heemskerk, Idse

doi:10.1101/2024.03.21.585753

Cited by 3 publications

(1 citation statement)

References 42 publications

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“…To differentiate cells for 2D gastruloids by micropatterning, we followed the protocol in 41 . Briefly, cells were resuspended in a single-cell suspension and seeded in laminin-coated micropatterned wells in mTeSR with ROCK inhibitor (RI) (MeChemExpress, cat# HY-10583).…”

Section: Data Availabilitymentioning

confidence: 99%

Endogenous FGFs drive ERK-dependent cell fate patterning in 2D human gastruloids

Jo,

Liu,

Patel

et al. 2024

Preprint

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The role of FGF is the least understood of the morphogens driving mammalian gastrulation. Here we investigated the function of FGF in a stem cell model for human gastrulation known as a 2D gastruloid. We found a ring of FGF-dependent ERK activity that closely follows the emergence of primitive streak (PS)-like cells but expands further inward. We showed that this ERK activity pattern is required for PS-like differentiation and that loss of PS-like cells upon FGF receptor inhibition can be rescued by directly activating ERK. We further demonstrated that the ERK-ring depends on localized activation of basally localized FGF receptors (FGFR) by endogenous FGF gradients. We confirm and extend previous studies in analyzing expression of FGF pathway components, showing the main receptor to be FGFR1 and the key ligands FGF2/4/17, similar to the human and monkey embryo but different from the mouse. In situ hybridization and scRNA-seq revealed thatFGF4andFGF17expression colocalize with the PS marker TBXT but onlyFGF17is maintained in nascent mesoderm and endoderm. FGF4 and FGF17 reduction both reduced ERK activity and differentiation to PS-like cells and their derivatives, indicating overlapping function. Thus, we have identified a previously unknown role for FGF-dependent ERK signaling in 2D gastruloids and possibly the human embryo, driven by a mechanism where FGF4 and FGF17 signal through basally localized FGFR1 to induce PS-like cells.

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Section: Data Availabilitymentioning

confidence: 99%

Endogenous FGFs drive ERK-dependent cell fate patterning in 2D human gastruloids

Jo,

Liu,

Patel

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

Endogenous FGFs drive ERK-dependent cell fate patterning in 2D human gastruloids

Jo,

Liu,

Patel

et al. 2024

Preprint

View full text Add to dashboard Cite

The role of FGF is the least understood of the morphogens driving mammalian gastrulation. Here we investigated the function of FGF in a stem cell model for human gastrulation known as a 2D gastruloid. We found a ring of FGF-dependent ERK activity that closely follows the emergence of primitive streak (PS)-like cells but expands further inward. We showed that this ERK activity pattern is required for PS-like differentiation and that loss of PS-like cells upon FGF receptor inhibition can be rescued by directly activating ERK. We further demonstrated that the ERK-ring depends on localized activation of basally localized FGF receptors (FGFR) by endogenous FGF gradients. We confirm and extend previous studies in analyzing expression of FGF pathway components, showing the main receptor to be FGFR1 and the key ligands FGF2/4/17, similar to the human and monkey embryo but different from the mouse. In situ hybridization and scRNA-seq revealed that FGF4 and FGF17 expression colocalize with the PS marker TBXT but only FGF17 is maintained in nascent mesoderm and endoderm. FGF4 and FGF17 reduction both reduced ERK activity and differentiation to PS-like cells and their derivatives, indicating overlapping function. Thus, we have identified a previously unknown role for FGF-dependent ERK signaling in 2D gastruloids and possibly the human embryo, driven by a mechanism where FGF4 and FGF17 signal through basally localized FGFR1 to induce PS-like cells.

show abstract

Endogenous FGFs drive ERK-dependent cell fate patterning in 2D human gastruloids

Jo,

Liu,

Patel

et al. 2024

Preprint

View full text Add to dashboard Cite

The role of FGF is the least understood of the morphogens driving mammalian gastrulation. Here we investigated the function of FGF in a stem cell model for human gastrulation known as a 2D gastruloid. We found a ring of FGF-dependent ERK activity that closely follows the emergence of primitive streak (PS)-like cells but expands further inward. We showed that this ERK activity pattern is required for PS-like differentiation and that loss of PS-like cells upon FGF receptor inhibition can be rescued by directly activating ERK. We further demonstrated that the ERK-ring depends on localized activation of basally localized FGF receptors (FGFR) by endogenous FGF gradients. We confirm and extend previous studies in analyzing expression of FGF pathway components, showing the main receptor to be FGFR1 and the key ligands FGF2/4/17, similar to the human and monkey embryo but different from the mouse. In situ hybridization and scRNA-seq revealed that FGF4 and FGF17 expression colocalize with the PS marker TBXT but only FGF17 is maintained in nascent mesoderm and endoderm. FGF4 and FGF17 reduction both reduced ERK activity and differentiation to PS-like cells and their derivatives, indicating overlapping function. Thus, we have identified a previously unknown role for FGF-dependent ERK signaling in 2D gastruloids and possibly the human embryo, driven by a mechanism where FGF4 and FGF17 signal through basally localized FGFR1 to induce PS-like cells.

show abstract

Extended culture of 2D gastruloids to model human mesoderm development

Cited by 3 publications

References 42 publications

Endogenous FGFs drive ERK-dependent cell fate patterning in 2D human gastruloids

Endogenous FGFs drive ERK-dependent cell fate patterning in 2D human gastruloids

Endogenous FGFs drive ERK-dependent cell fate patterning in 2D human gastruloids

Endogenous FGFs drive ERK-dependent cell fate patterning in 2D human gastruloids

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