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Microfabrication-based modulation of embryonic stem cell differentiation.

Jaesung Park1, Cheul H Cho, Natesh Parashurama

  • 1Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Shriners Hospitals for Children and Harvard Medical School, Boston, Massachusetts 02114, USA.

Lab on a Chip
|July 27, 2007
PubMed
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Controlled differentiation of embryonic stem cells (ES cells) is crucial. Uniform aggregate size, achieved via microfabricated stencils, significantly impacts ES cell differentiation into specific germ layers.

Area of Science:

  • Stem cell biology
  • Developmental biology
  • Biomaterials engineering

Background:

  • Embryonic stem (ES) cells aggregate spontaneously during differentiation, with cell-cell communication influencing the process.
  • Current methods for producing ES cell aggregates lack precise size control, hindering controlled differentiation.
  • Uniform aggregate size is essential for reproducible ES cell differentiation studies.

Purpose of the Study:

  • To develop a microfabrication technique for producing uniform mouse ES (mES) cell aggregates of controlled sizes.
  • To investigate the impact of initial mES cell aggregate size on differentiation into the three germ layers.
  • To establish a reliable method for size-controlled ES cell aggregate formation for differentiation studies.

Main Methods:

Related Experiment Videos

  • Microfabrication of adhesive stencils to create mES cell aggregates ranging from 100 to 500 micrometers in diameter.
  • Induction of differentiation for 20 days.
  • Analysis of stem cell populations using gene and protein expression assays and biochemical functions.

    • Main Results:

    • Germ layer differentiation is dependent on the initial size of the mES cell aggregate.
    • Aggregates of 300 micrometers showed differentiation profiles similar to the "hanging drop" method.
    • Smaller aggregates (100 micrometers) favored ectodermal differentiation, while larger aggregates (500 micrometers) favored mesodermal and endodermal differentiation.

    Conclusions:

    • Initial aggregate size is a critical factor influencing ES cell differentiation outcomes, including germ layer selection.
    • Microfabricated stencils provide a reliable method for generating size-controlled ES cell aggregates.
    • This technique enables precise studies on how aggregate size affects stem cell differentiation and developmental potential.