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Reproductive Sciences
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Spindle and Chromosome Changes of Human MII Oocytes During Incubation After Slow Freezing/Fast Thawing Procedures

Shanshan Gao, MD

Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, Jinan, 250021, China, Key Laboratory of Reproductive Medicine, Shandong Province, China

Yuan Li, MD, PhD

Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, Jinan, 250021, China, Key Laboratory of Reproductive Medicine, Shandong Province, China

Xuan Gao, MD

Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, Jinan, 250021, China, Key Laboratory of Reproductive Medicine, Shandong Province, China

Jingmei Hu, MD

Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, Jinan, 250021, China, Key Laboratory of Reproductive Medicine, Shandong Province, China

Huijun Yang, MD

Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, Jinan, 250021, China, Key Laboratory of Reproductive Medicine, Shandong Province, China

Zi-Jiang Chen, MD, PhD

Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, Jinan, 250021, China, zjchen59{at}126.com, Key Laboratory of Reproductive Medicine, Shandong Province, China

This experiment investigated the optimal time required for cryopreserved human oocytes to reform their spindles upon re-warming. Metaphase II oocytes were cryopreserved with a slow freezing method. Oocytes from each patient were randomly allocated into one of the 3 groups with different culture periods: 1, 3, or 5 hours, respectively, after thawing. Tubulin and chromosome configurations were visualized by confocal microscopy after immunostainings. By morphological assessment, 87.3% oocytes survived the process of freezing and thawing. Oocytes with normal spindle configuration increased significantly after 3 or 5 hours of incubation compared to those incubated for only 1 hour ( P < 0.05). There were no differences in the chromosome configurations among the treatment groups ( P > 0.05). This experiment demonstrated that cryopreserved human oocytes need a certain minimum period of incubation time (3 h) to recover their disrupted MII spindles and this information can be used in development of human IVF protocols with frozen oocytes.

Key Words: Oocyte • freezing • thawing • chromosome • meiotic spindle.

This version was published on April 1, 2009

Reproductive Sciences, Vol. 16, No. 4, 391-396 (2009)
DOI: 10.1177/1933719108327590
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