The oocyte spindle is preserved by 1,2-propanediol during slow freezing

Chang, Ching-Chien; Sung, Li-Ying; Lin, Chih-Jen; Kort, Hilton I.; Yang, Xiangzhong; Tian, X. Cindy; Nagy, Zsolt Peter

doi:10.1016/j.fertnstert.2009.01.106

« Previous Next »Fertility and Sterility
Volume 93, Issue 5 , Pages 1430-1439, 15 March 2010

The oocyte spindle is preserved by 1,2-propanediol during slow freezing

Presented in part at the annual meeting of American Society for Reproductive Medicine, Washington, DC, October13–17, 2007.

Ching-Chien Chang, Ph.D.
- Reproductive Biology Associates, Atlanta, Georgia
Li-Ying Sung, Ph.D.
- Center for Regenerative Biology, University of Connecticut, Storrs, Connecticut
- Department of Animal Science, University of Connecticut, Storrs, Connecticut
Chih-Jen Lin, M.Sc.
- Center for Regenerative Biology, University of Connecticut, Storrs, Connecticut
- Department of Animal Science, University of Connecticut, Storrs, Connecticut
Hilton I. Kort, M.D.
- Reproductive Biology Associates, Atlanta, Georgia
Xiangzhong Yang, Ph.D.
- Center for Regenerative Biology, University of Connecticut, Storrs, Connecticut
- Department of Animal Science, University of Connecticut, Storrs, Connecticut
X. Cindy Tian, Ph.D.
- Center for Regenerative Biology, University of Connecticut, Storrs, Connecticut
- Department of Animal Science, University of Connecticut, Storrs, Connecticut
Zsolt Peter Nagy, M.D., Ph.D.
- Reproductive Biology Associates, Atlanta, Georgia
- Reprint requests: Zsolt Peter Nagy, M.D., Ph.D., Reproductive Biology Associates, 1150 Lake Hearn Drive, Suite 600, Atlanta, GA 30342 (FAX: 404-256-1912).

Received 24 October 2008; received in revised form 10 December 2008; accepted 9 January 2009. published online 26 March 2009.

Objective

To investigate the specific changes in oocyte spindle subjected to severe challenges of low temperature, as well as to examine the effect of cryoprotectants in preserving oocyte spindle during cryopreservation.

Design

In vitro experimental study.

Setting

Academic research laboratory.

Animal(s)

B6D2F1 (C57BL/6 X DBA/2) mice.

Intervention(s)

Mouse oocytes were cryopreserved using a slow freezing method in a sodium-depleted medium with 1.5 mol/l 1,2-propanediol (PROH) and 0.3 M sucrose. To examine the spindle, oocytes were fixed before, during, and after cryopreservation, and oocytes were analyzed by immunocytochemistry and confocal microscopy.

Result(s)

The MII spindle was preserved during the slow freezing, because the cryoprotectant PROH was found to support the organization of MII spindle in resisting the subzero temperature. In contrast, the MII spindle was disassembled gradually during the thawing process with or without PROH. Most of the oocytes were able to recover the MII spindle after thawing, but a portion of thawed oocytes could not sustain the meiotic spindle because of parthenogenetic activation.

Conclusion(s)

1,2-Propanediol can support the organization of MII spindle to defy the subphysiologic temperature; however, the PROH cannot sustain oocyte spindle structure after the subsequent thawing process.

Key Words: Oocyte cryopreservation, meiotic spindle, 1,2-propanediol

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C.-C.C. had nothing to disclose. L.-Y.S. had nothing to disclose. C.-J.L. has nothing to disclose. H.I.K. has nothing to disclose. X.Y. had nothing to disclose. X.C.T. had nothing to disclose. Z.P. N. has nothing to disclose.

PII: S0015-0282(09)00144-7

doi:10.1016/j.fertnstert.2009.01.106

« Previous Next »Fertility and Sterility
Volume 93, Issue 5 , Pages 1430-1439, 15 March 2010

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