The effect of osmotic stress on the cell volume, metaphase II spindle and developmental potential of in vitro matured porcine oocytes.
Porcine animal models are used to advance our understanding of human physiology. Current research is also directed at methods to produce transgenic pigs. Cryobanking gametes and embryos can facilitate the preservation of valuable genotypes, yet cryopreserving oocytes from pigs has proven very challenging. The current study was designed to understand the effects of anisotonic solutions on in vitro matured porcine oocytes as a first step toward designing improved cryopreservation procedures. We hypothesized that the proportion of oocytes demonstrating a normal spindle apparatus and in vitro developmental potential would be proportional to the solution osmolality. Oocytes were incubated for 10 min at 38 degrees C in various hypo- or hypertonic solutions, and an isotonic control solution and then assessed for these two parameters. Our results support the hypothesis, with an increasing proportion of spindles showing a disrupted structure as the levels of anisotonic exposure diverge from isotonic. Only about half of the oocytes maintained developmental potential after exposure to anisotonic solutions compared to untreated controls. Oocyte volume displayed a linear response to anisotonic solutions as expected, with an estimated relative osmotically inactive cell volume of 0.178. The results from this study provide initial biophysical data to characterize porcine oocytes. The results from future experiments designed to determine the membrane permeability to various cryoprotectants will allow predictive modeling of optimal cryopreservation parameters and provide a basis for designing improved cryopreservation procedures.
Mullen
S F
SF
The Comparative Medicine Center and Department of Veterinary Pathobiology, The University of Missouri at Columbia, Columbia, MO 65211, USA.
Rosenbaum
M
M
Critser
J K
JK
D000818:Animals; D001755:Blastocyst / Q000502:physiology / Q000648:ultrastructure; D048429:Cell Size; D015925:Cryopreservation / Q000379:methods; D005260:Female; D005307:Fertilization in Vitro; D008677:Metaphase; D009865:Oocytes / Q000502:physiology* / Q000648:ultrastructure; D009994:Osmolar Concentration; D009997:Osmotic Pressure; D008941:Spindle Apparatus / Q000648:ultrastructure*; D013552:Swine