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Published on: June 21, 2014
Skeletal myogenesis: the preferred pathway of chick embryo epiblast cells in vitro
M George-Weinstein1, J Gerhart, R Reed
1Department of Anatomy, Philadelphia College of Osteopathic Medicine, Pennsylvania 19131, USA.
Insights
Chick embryo epiblast cells can form skeletal muscle early in development. Cell and tissue interactions in the embryo normally prevent this muscle differentiation.
Area of Science:
- Developmental biology
- Cell biology
- Embryology
Background:
- The epiblast is the precursor to all embryonic tissues in chick embryos.
- Understanding early cell differentiation is crucial for developmental biology.
Purpose of the Study:
- To investigate the potential of epiblast cells to form skeletal muscle before entering the primitive streak.
- To determine the role of cell and tissue interactions in regulating myogenesis.
Main Methods:
- Isolation and dissociation of epiblast cells from chick embryos.
- In vitro culture of single-cell suspensions at high density.
- Analysis of myogenesis and MyoD messenger RNA expression.
Main Results:
- Epiblast cells readily differentiated into skeletal muscle in vitro, with myogenesis beginning on day one.
- MyoD messenger RNA was present and translated in cultured epiblast cells.
- Cells from regions not typically forming muscle in vivo successfully differentiated into muscle in vitro.
- Intact epiblasts or those cultured with mesoderm/hypoblast did not undergo myogenesis.
Conclusions:
- Myogenic potential is widespread in the primitive streak stage epiblast.
- Muscle differentiation in epiblast cells occurs relatively autonomously in culture.
- Embryonic cell and tissue interactions can inhibit muscle differentiation.
Abstract:
The epiblast layer of the chick embryo gives rise to all embryonic tissues. In vitro analyses were carried out to determine whether epiblast cells could form skeletal muscle prior to entry into the primitive streak. Epiblasts were separated from the mesoderm, hypoblast, and primitive streak, dissociated to produce a single cell suspension, and plated at high density. Myogenesis began on the first day in culture, and by the fifth day most cells had differentiated into skeletal muscle. Some cells differentiated without replicating. MyoD messenger RNA was present in epiblast tissue and translated in practically all cells in culture. Cells from regions of the epiblast which do not form muscle later in the embryo did so in vitro. Epiblasts cultured for 2 days as an intact epithelium, or in the presence of the mesoderm and hypoblast, did not undergo myogenesis. These findings demonstrate that myogenic potential is wide-spread within the primitive streak stage epiblast, and that muscle differentiation, which occurs relatively autonomously in culture, can be prevented by cell and tissue interactions.
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