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Updated: Jul 17, 2026
Multi-Photon Laser Ablation of Cytoplasmic Microtubule Organizing Centers in Mouse Oocytes
Published on: November 11, 2022
Reconstruction of ooplasm recipient oocytes with frozen-thawed donor microcytoplasts and influence on the
Anuradha P Goud1, Pravin T Goud, Michael P Diamond
1Department of Obstetrics and Gynecology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA. agoud@med.wayne.edu
Objective(S):
To cryopreserve micromanipulated ooplast segments (microcytoplasts) from mouse oocytes, compare microcytoplast and parent or recipient oocyte fusion performed within or without the zona pellucida, compare electrofusion of fresh or frozen oocyte with frozen-thawed microcytoplasts, and assess spindle integrity after reconstruction of oocytes.
Design:
Prospective experimental study.
Setting:
University-based experimental laboratory.
Animal(S):
Mouse (MII) oocytes obtained after superovulation (n = 363).
Intervention(S):
Micromanipulation of oocytes (n = 363) into microcytoplasts (n = 181), cryopreservation of microcytoplasts along with parent and sibling control oocytes (n = 182), reconstruction by electrofusion of microcytoplast and parent or recipient oocyte performed with (group A, n = 35) or without a zona pellucida (group B, n = 32), comparison of electrofusion of fresh oocyte (group C, n = 40) or frozen oocyte (group D, n = 36) with frozen-thawed microcytoplasts fused within zona, and assessment of spindle morphology of reconstructed oocyte.
Main Outcome Measure(S):
Post-thaw survival, success of fusion, and spindle integrity as assessed by immunostaining.
Result(S):
Higher success of post-thaw fusion was seen in group A (91.4%) compared with group B (56.2%). The post-thaw fusion of microcytoplasts with either fresh or frozen oocytes was not significantly different. Spindle integrity was 82.5% in group C as compared with 47.2% in group D.
Conclusion(S):
Microcytoplasts created from oocytes can be successfully cryopreserved, thawed, and used to reconstruct oocytes with intact spindles.
Insights
Cryopreserved mouse ooplast segments (microcytoplasts) can be successfully used to reconstruct oocytes. This method preserves spindle integrity, offering a promising approach for assisted reproduction technologies.
Area of Science:
- Reproductive Biology
- Cryobiology
- Assisted Reproductive Technologies
Background:
- Oocyte cryopreservation is crucial for assisted reproduction.
- Micromanipulation techniques allow for the isolation of ooplast segments, termed microcytoplasts.
- The viability and functionality of these microcytoplasts post-cryopreservation require thorough investigation.
Purpose of the Study:
- To establish a cryopreservation protocol for mouse ooplast segments (microcytoplasts).
- To compare the efficiency of oocyte reconstruction using microcytoplasts with or without the zona pellucida.
- To evaluate the spindle integrity of reconstructed oocytes following fusion with fresh or frozen-thawed microcytoplasts.
Main Methods:
- Mouse oocytes underwent micromanipulation to create microcytoplasts.
- Microcytoplasts, along with control oocytes, were cryopreserved and thawed.
- Reconstruction involved electrofusion of microcytoplasts with parent or recipient oocytes, both within and without the zona pellucida.
- Spindle morphology was assessed using immunostaining.
Main Results:
- Fusion success rates were significantly higher when performed within the zona pellucida (91.4%) compared to without (56.2%).
- Post-thaw fusion efficiency did not differ significantly between fresh and frozen oocytes when fused with microcytoplasts.
- Spindle integrity was notably preserved in reconstructed oocytes (82.5%) compared to those with compromised spindles (47.2%).
Conclusions:
- Cryopreservation of microcytoplasts is feasible and effective for oocyte reconstruction.
- Reconstructed oocytes utilizing cryopreserved microcytoplasts can maintain spindle integrity.
- This technique holds potential for advancing fertility preservation and reproductive medicine.
