Journal of Assisted Reproduction and Genetics

, Volume 10, Issue 6, pp 427–433 | Cite as

Trophectoderm microbiopsy in murine blastocysts: Comparison of four methods

  • Sandra A. Carson
  • William L. Gentry
  • Albert L. Smith
  • John E. Buster
Animal Investigations

Purpose

This study compares four trophectoderm microbiopsy techniques for removal of blastomeres from murine blastocysts: (1) aspiration,trophectoderm pipetted through the zona; (2) incision,trophectoderm excised with a microrazor; (3) slit/excision,the zona slit and herniating trophectoderm excised; and (4) hatch/excision,trophectoderm cells excised after spontaneous hatching.

Results

Murine blastocysts were comparatively biopsied using one of four methods and contrasted to zona slit and nonmicromanipulated controls. Operative cellular injury was assessed by uptake of fluorescein isothiocyanate (FITC). Postoperative embryonic viability was assessed by blastocoele reexpansion and hatching inner cell mass development and trophectoderm plating. All techniques yielded cells available for genetic analysis.

Conclusions

The slit/excision technique and hatch/excision techniques exhibited lower operative injury and the higher postoperative viability than aspiration or incision. The slit/excision and the hatch/excision techniques, though requiring two operative steps, appear to be the least damaging of the four methods. Therefore, they should be most applicable to human blastocysts obtained either by extended culture in vitro or by uterine lavage.

Key words

blastocysts blastomere biopsy murine human preimplantation diagnosis 

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Copyright information

© Plenum Publishing Corporation 1993

Authors and Affiliations

  • Sandra A. Carson
    • 1
  • William L. Gentry
    • 2
  • Albert L. Smith
    • 3
  • John E. Buster
    • 1
  1. 1.Department of Obstetrics and GynecologyThe University of Tennessee, Memphis, Division of Reproductive EndocrinologyMemphisUSA
  2. 2.IndianapolisUSA
  3. 3.Advanced Fertility and Reproductive Endocrinology AssociatesSan AntonioUSA

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