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1 January 2003 Artificial Expression of Aquaporin-3 Improves the Survival of Mouse Oocytes after Cryopreservation
Keisuke Edashige, Yohei Yamaji, F. W. Kleinhans, Magosaburo Kasai
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Successful cryopreservation of mammalian cells requires rapid transport of water and cryoprotective solutes across the plasma membrane. Aquaporin-3 is known as a water/solute channel that can transport water and neutral solutes such as glycerol. In this study we examined whether artificial expression of aquaporin-3 in mouse oocytes can improve water and glycerol permeability and oocyte survival after cryopreservation. Immature mouse oocytes were injected with aquaporin-3 cRNA and were cultured for 12 h. Then the hydraulic conductivity (LP) and glycerol permeability (PGLY) of matured oocytes were determined from the relative volume changes in 10% glycerol in PB1 medium at 25°C. Mean ± SD values of LP and PGLY of cRNA-injected oocytes (3.09 ± 1.22 μm min−1 atm−1 and 3.69 ± 1.47 × 10−3 cm/min, respectively; numbers of oocytes = 25) were significantly higher than those of noninjected oocytes (0.83 ± 0.02 μm min−1 atm−1 and 0.07 ± 0.02 × 10−3 cm/min, respectively; n = 13) and water-injected oocytes (0.87 ± 0.10 μm min−1 atm−1 and 0.08 ± 0.02 × 10−3 cm/min, respectively; n = 20). After cryopreservation in a glycerol-based solution, 74% of cRNA-injected oocytes (n = 27) survived as assessed by their morphological appearance, whereas none of the water-injected oocytes survived (n = 10). When cRNA-injected oocytes that survived cryopreservation were inseminated in vitro, the penetration rate was 40% (n = 48) and the cleavage rate was 31% (n = 70), showing that oocytes retain their ability to be fertilized. This is the first report to show that artificial expression of a water/solute channel in a cell improves its survival after cryopreservation. This approach may enable cryopreservation of cells that have been difficult to cryopreserve.

Keisuke Edashige, Yohei Yamaji, F. W. Kleinhans, and Magosaburo Kasai "Artificial Expression of Aquaporin-3 Improves the Survival of Mouse Oocytes after Cryopreservation," Biology of Reproduction 68(1), 87-94, (1 January 2003).
Received: 7 December 2001; Accepted: 1 July 2002; Published: 1 January 2003
oocyte development
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