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Bovine embryonic stem cells.

Marsha Roach1, Li Wang, Xiangzhong Yang

  • 1Pfizer Global Research and Development, Genetically Modified Models CoE, Groton, Connecticut, USA.

Methods in Enzymology
|December 5, 2006
PubMed
Summary
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This study details methods for isolating and maintaining bovine embryonic stem (bES) cells, confirming their pluripotency through marker expression and differentiation. These bES cells resemble mouse and human counterparts, offering potential for patient-specific cell therapies.

Area of Science:

  • Veterinary Science
  • Stem Cell Biology
  • Reproductive Biology

Background:

  • Bovine embryonic stem (bES) cell lines exhibit variable morphology and marker expression, challenging their characterization.
  • Key markers for pluripotency, including alkaline phosphatase (ALPL), SSEA4, and OCT4, are inconsistently reported in existing bES cell lines.

Purpose of the Study:

  • To introduce reliable methods for isolating and maintaining bovine ES cells.
  • To characterize bES cell lines based on morphology and pluripotency marker expression.
  • To explore alternative methods for generating blastocysts for potential patient-specific ES cell generation.

Main Methods:

  • Isolation and culture of bovine ES cells.
  • Assessment of pluripotency markers (ALPL, SSEA4, OCT4) via staining.

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  • Evaluation of differentiation potential into three germ layers.
  • Generation of blastocysts using in vitro fertilization (IVF), parthenogenetic activation, and somatic cell nuclear transfer (SCNT).

    • Main Results:

    • Bovine ES cells were cultured in large colonies, exhibiting morphology similar to mouse/human ES and EG cells.
    • Cells consistently stained positive for ALPL, SSEA4, and OCT4, mirroring blastocyst patterns.
    • These bES cells maintained pluripotency and differentiation capacity for over a year.
    • Alternative methods for blastocyst generation were successfully introduced.

    Conclusions:

    • Established bES cell lines demonstrate consistent pluripotency markers and differentiation capabilities.
    • The described methods provide a robust approach for bovine ES cell culture.
    • Alternative blastocyst generation techniques offer pathways for patient-specific stem cell applications.