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Cell Biology JoVE (Open Access) Citable · DOI

Mouse Oocyte Microinjection, Maturation and Ploidy Assessment

DOI: 10.3791/2851-v
What you'll learn
  • ✓ Perform microinjection of materials into mouse oocytes
  • ✓ Assess meiotic maturation and ploidy status in eggs
  • ✓ Evaluate chromosome segregation errors and aneuploidy
Protocol

Oocytes are prone to aneuploidy due to errors in chromosome segregation during meiotic maturation. Aneuploid eggs can cause infertility, miscarriages or developmental disorders like Down syndrome. Here, we describe methods to introduce materials of choice into oocytes and methods to study meiotic maturation and assess ploidy.

Difficulty
advanced
Total time
~2–3 hours per experimental cohort (collection through ploidy analysis)
Model organism
Mouse (strain unspecified)
Biosafety
BSL-1

Steps

1
Collect mouse oocytes from ovarian follicles

Isolate metaphase II oocytes from mouse ovaries using standard collection techniques. Prepare oocytes for downstream microinjection and maturation studies.

▶ 01:00
2
Microinject oocytes and induce meiotic maturation

Introduce experimental materials into oocytes via microinjection. Culture injected oocytes to permit meiotic maturation and completion of meiosis I/II.

▶ 02:28
3
Perform ploidy analysis on mature eggs

Assess chromosome number and ploidy status in matured oocytes using cytogenetic or molecular methods. Identify euploid versus aneuploid eggs.

▶ 04:14
4
Interpret euploid metaphase II egg profiles

Evaluate morphological and chromosomal characteristics of normal euploid metaphase II eggs as positive controls and benchmarks for ploidy assessment.

▶ 06:04
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