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Related Concept Videos

Determination01:51

Determination

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During embryogenesis, cells become progressively committed to different fates through a two-step process: specification followed by determination. Specification is demonstrated by removing a segment of an early embryo, “neutrally” culturing the tissue in vitro—for example, in a petri dish with simple medium—and then observing the derivatives. If the cultured region gives rise to cell types that it would normally generate in the embryo, this means that it is specified. In...
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Rapid and Efficient Generation of Neurons from Human Pluripotent Stem Cells in a Multititre Plate Format
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Deterministic HOX patterning in human pluripotent stem cell-derived neuroectoderm.

Ethan S Lippmann1, Clay E Williams2, David A Ruhl3

  • 1Department of Biomedical Engineering, University of Wisconsin, Madison, WI 53706, USA; Wisconsin Institute for Discovery, University of Wisconsin, Madison, WI 53706, USA.

Stem Cell Reports
|April 7, 2015
PubMed
Summary
This summary is machine-generated.

This study details a new method for precisely patterning human pluripotent stem cells (hPSCs) into specific hindbrain and spinal cord neural cells. The approach ensures accurate HOX gene activation for regional neural fate determination.

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Area of Science:

  • Developmental Biology
  • Stem Cell Biology
  • Neuroscience

Background:

  • HOX gene expression patterns neural development in vivo, assigning regional identities to the hindbrain and spinal cord.
  • Current in vitro methods for differentiating human pluripotent stem cells (hPSCs) into posterior neural fates lack precision, resulting in broadly specified cell populations.

Purpose of the Study:

  • To develop a defined protocol for precise regional patterning of hPSCs towards hindbrain and spinal cord neural fates.
  • To achieve controlled, collinear HOX gene activation during hPSC differentiation for predictable neural phenotype generation.

Main Methods:

  • Sequential activation of fibroblast growth factor, Wnt/β-catenin, and growth differentiation factor signaling in hPSCs.
  • Introduction of retinoic acid at specific time points to halt HOX activation and induce neuroectoderm formation.
  • Characterization of SOX2, Brachyury, and PAX6 expression to define neuromesoderm and neuroectoderm stages.

Main Results:

  • A 7-day differentiation process yielded stable, homogenous SOX2(+)/Brachyury(+) neuromesoderm with progressive, collinear HOX activation.
  • Retinoic acid treatment at any stage arrested HOX activation, generating SOX2(+)/PAX6(+) neuroectoderm with discrete HOX profiles.
  • Generated cells could be further differentiated into region-specific cell types, such as motor neurons.

Conclusions:

  • This defined protocol enables precise control over HOX gene expression during hPSC differentiation.
  • The method significantly enhances the ability to derive region-specific neural cells from the hindbrain and spinal cord.
  • This approach expands the toolkit for generating diverse neural phenotypes for research and therapeutic applications.