| Experiments were conducted to study the effects of Rhodiola Sachalinensis Polysaccaride (RSP) on developmental competence of porcine oocytes obtained from abattoir ovaries following in vitro maturation and cryopreservation. The results showed that:(1) Firstly porcine oocytes derived from follicle of different sizes were allocated to three treatments according follicles size:>6mm,2-6mm and<2mm. The nuclear maturation rates of secondary follicles (2-6mm)(83.90±1.54%)were higher than that of competent follicles(>6mm)(80.90±2.69%, P>0.5), but both of them were significantly higher than that of small follicles s(>6mm)(47.03±4.72%,P<0.05).(2) The effect of different volum mature culture on porcine oocyte in vitro maturation was investigated.15,35,65porcine oocytes were matured in50μL,250μL,500μL mature culture. The results showed that the the nuclear maturation rates using250uL(79.53±3.48%) and500uL(79.71±5.91%) culture medium were significantly higher than that culture in50uLdrop(65.41±6.28%,P<0.05), and were not significantly different between them.(3) The effect of Rhodiola Sachalinensis Polysaccaride (RSP) on developmental competence of porcine oocytes. The nuclear maturation rates in each RSP treatment were higher than that of control group (83.50±1.00%,P>0.05); The GV stage oocytes treated with6and60mg·L-1RSP in the vitro maturation medium could significantly increase the GSH content (P<0.05), and decrease the ROS level (P>0.05); After PA, the cleavage rate of60mg·L-1RSP (88.44±5.06%) and the blastocyst rate of6mg·L-RSP (40.94±8.23%) were the highest, and were significantly higher than those of control group (72.16±7.38%,24.06±5.16%,P<0.05); IVF cleavage rate in each RSP treatment was significantly increased (P<0.05), IVF blastocyst rate (19.20±3.13%) derived from oocytes treated with60mg·L-1RSP was significantly higher than that of control group (9.60±0.65%,P<0.05). (4) The effect of self-made carries (Hemi-straw), OPS and straw on the vitrification of porcine MII oocytes was investigated. The results showed that the morphologically normal rates and survival rates which were obtained from oocytes vitrified-warmed by using Hemi-straw(87.36±1.93%,73.78±4.06%) were higher than using OPS(84.37±3.49%,70.48±1.06%,P>0.05),but were significantly higher than using staw(67.43±2.72%,54.40±3.80,P<0.05).(5) In order to improve the vitrification effect of porcine matured oocytes, this experiment was designed to use the Vit-Master vitrification machine to freeze the porcine in vitro matured oocytes. The result showed that the cooling rate (21345±1768℃/min) of slush liquid nitrogen from Vit-Master were higher than that of normal liquid nitrogen (11982±1936℃/min). There were no significant differences in all the groups in morphologically normal rate (P>0.05). The survival rates of the groups from Vit-Master groups (A’(57.87±10.66%,), B’(76.5±10.33%), C’(79.07±14.90%), and D’(81.94±6.98%))was higher than those of A, B, C and D groups without Vit-Master treatment (49.73±2.91%,60.63±12.45%,71.33±14.38%and78.50±7.40%,P>0.05). The survival rate of groups of B’and C’ were much higher than the groups of C and D (P>0.05), but the groups of A and A’were significantly lower than the groups of B’、C’and D (P<0.05). It was concluded that the use of Vit-Master in the vitrification could increase the cooling rate and reduce the concentration of CPAs, then improve the post-thawed oocyte survival rate.(6) The effect of Rhodiola Sachalinensis Polysaccaride (RSP) on vitrification of porcine oocytes. Immature oocytes were matured in vitro and supplemented with different concentrations of RSP, then the oocytes matured for44h were carried out vitrification. The result showed that the survival rates and the cleavage rates in each RSP treatment were higher than that of control group (72.44±2.64%,4.33±1.11%, P>0.05). The cleavage rate (8.68±2.08%) derived from oocytes treated with60mg·L-RSP was significantly higher than that of control group. It was supplemented with different concentrations of RSP in pretreatment and freezing solution. There were no significant difference between grops, either the survival rates or the cleavage rates. |
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