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Related Experiment Videos

Selection of genetically modified chicken blastodermal cells by magnetic-activated cell sorting.

Q Wei1, B A Croy, R J Etches

  • 1Department of Biomedical Sciences, University of Guelph, Ontario, Canada. wqingxia@hotmail.com

Poultry Science
|January 5, 2002
PubMed
Summary
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Magnetic-activated cell sorting (MACS) enriches genetically modified chicken blastodermal cells (CBC). This method improves the efficiency of producing transgenic chickens by increasing the proportion of modified CBC for embryo manipulation.

Area of Science:

  • Animal Biotechnology
  • Developmental Biology
  • Cell Biology

Background:

  • Production of transgenic chickens relies on genetic modification of chicken blastodermal cells (CBC).
  • Higher proportions of stably transfected CBC in embryo inoculum correlate with increased chimeric bird frequency.
  • Enrichment of transfected CBC is crucial for efficient generation of transgenic poultry.

Purpose of the Study:

  • To evaluate magnetic-activated cell sorting (MACS) as a method for enriching transfected CBC.
  • To assess the effectiveness of MACS in isolating genetically modified chicken cells.
  • To determine if magnetically sorted CBC retain their developmental potential.

Main Methods:

  • Chicken blastodermal cells were coelectroporated with pmiwZ and pMACS Kk plasmids.

Related Experiment Videos

  • Cells were sorted using magnetic-activated cell sorting (MACS) based on mouse H-2Kk surface molecule expression.
  • Transfection efficiency was assessed by X-gal staining for lacZ expression.

    • Main Results:

    • MACS resulted in a 1.4-fold enrichment of lacZ-positive CBC after 48 hours of culture.
    • The enriched cell population contributed to both extra-embryonic and intra-embryonic tissues in recipient embryos.
    • A marginal increase in intra-embryonic contribution was observed from the magnetically sorted cells.

    Conclusions:

    • Magnetic-activated cell sorting (MACS) effectively enriches genetically modified chicken blastodermal cells (CBC).
    • Magnetically sorted CBC maintain their developmental potential, contributing to ectodermal and mesodermal lineages.
    • This technique shows promise for enhancing the production of transgenic birds through improved genetic modification efficiency.