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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

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.

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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.
  • 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.