Hum. Reprod. Advance Access originally published online on April 7, 2004
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Human Reproduction, Vol. 19, No. 5, 1148-1154,
May 2004
© 2004 European Society of Human Reproduction and Embryology
The effect of osmotic stress on the metaphase II spindle of human oocytes, and the relevance to cryopreservation.
1 Comparative Medicine Center and Department of Veterinary Pathobiology, University of Missouri at Columbia, Columbia MO 65211, and 2 The Atlanta Center for Reproductive Medicine, Woodstock GA 30189, USA
3 To whom correspondence should be addressed. e-mail: critserj{at}missouri.edu
BACKGROUND: Knowing osmotic tolerance limits is important in the design of optimal cryopreservation procedures for cells. METHODS: Mature human oocytes were exposed to anisosmotic sucrose solutions at concentrations of 35, 75, 150, 600, 1200, or 2400 (±5) milliosmolal (mOsm) at 37°C. A control treatment at 290 mOsm was also utilized. Oocytes were randomly allocated to each experimental treatment. After the treatment, the oocytes were cultured for 1 h, then fixed in cold methanol. Immunocytochemical staining and fluorescence microscopy were used to assess the morphology of the metaphase II (MII) spindle. Logistic regression was used to determine if media osmolality had a significant effect on spindle structure. RESULTS: Osmolality was a significant predictor of spindle morphology. Hyposmotic effects at 35, 75, and 150 mOsm resulted in 100, 67, and 56% of oocytes having abnormal spindles, respectively. Hyperosmotic effects at 600, 1200, and 2400 mOsm resulted in 44, 44, and 100% of the spindles with abnormal structure, respectively. CONCLUSIONS: Anisosmotic conditions lead to disruption of the MII spindle in human oocytes. Applying this fundamental knowledge to human oocyte cryopreservation should result in increased numbers of cells maintaining viability.
Key words: cryopreservation/human/MII spindle/oocytes/osmotic tolerance
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  Y.-L. Miao, K. Kikuchi, Q.-Y. Sun, and H. Schatten Oocyte aging: cellular and molecular changes, developmental potential and reversal possibility Hum. Reprod. Update, September 1, 2009; 15(5): 573 - 585. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Shanshan Gao, Yuan Li, Xuan Gao, Jingmei Hu, Huijun Yang, and Z.-J. Chen Spindle and Chromosome Changes of Human MII Oocytes During Incubation After Slow Freezing/Fast Thawing Procedures Reproductive Sciences, April 1, 2009; 16(4): 391 - 396. [Abstract] [PDF]
|
|
|
|
|
|
D. A. Gook and D. H. Edgar Human oocyte cryopreservation Hum. Reprod. Update, November 1, 2007; 13(6): 591 - 605. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. De Santis, G. Coticchio, S. Paynter, D. Albertini, K. Hutt, I. Cino, M. Iaccarino, A. Gambardella, C. Flamigni, and A. Borini Permeability of human oocytes to ethylene glycol and their survival and spindle configurations after slow cooling cryopreservation Hum. Reprod., October 1, 2007; 22(10): 2776 - 2783. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Van den Abbeel, U. Schneider, J. Liu, Y. Agca, J. K Critser, and A. Van Steirteghem Osmotic responses and tolerance limits to changes in external osmolalities, and oolemma permeability characteristics, of human in vitro matured MII oocytes Hum. Reprod., July 1, 2007; 22(7): 1959 - 1972. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.A. Nottola, G. Macchiarelli, G. Coticchio, S. Bianchi, S. Cecconi, L. De Santis, G. Scaravelli, C. Flamigni, and A. Borini Ultrastructure of human mature oocytes after slow cooling cryopreservation using different sucrose concentrations Hum. Reprod., April 1, 2007; 22(4): 1123 - 1133. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. LaRosa and S. M. Downs Stress Stimulates AMP-Activated Protein Kinase and Meiotic Resumption in Mouse Oocytes Biol Reprod, March 1, 2006; 74(3): 585 - 592. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Ghetler, S. Yavin, R. Shalgi, and A. Arav The effect of chilling on membrane lipid phase transition in human oocytes and zygotes Hum. Reprod., December 1, 2005; 20(12): 3385 - 3389. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.J. Paynter, A. Borini, V. Bianchi, L. De Santis, C. Flamigni, and G. Coticchio Volume changes of mature human oocytes on exposure to cryoprotectant solutions used in slow cooling procedures Hum. Reprod., May 1, 2005; 20(5): 1194 - 1199. [Abstract] [Full Text] [PDF]
|
|