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In this experiment, 44 New Zealand female rabbits as experimental material, research rabbit ovarian surface in vitro maturation of oocytes, in vitro fertilization, and the fertilized eggs of different in vitro culture conditions, in order to optimize the rabbit oocytes in vitro maturation, in vitro fertilization, and the fertilized egg culture the system, the results are as follows: 1. cutting method, acupuncture extrusion method to collect the ovarian surface oocytes to test the impact of different collection methods to collect the number of ovarian oocytes. The results show that: the needling extrusion process the obtained oocytes (23.63 / PCS), significantly higher than the cutting method (16.50 / PCS) (p LT; 0.05). Compared adding different concentrations of FSH, LH rabbit oocytes in vitro mature, the results show that: 1 U / mL FSH 2 U / mL LH maturity of the highest rates of 46.62%, which was significantly higher than (TCM199 10? the S)'s maturation rate (13 71%, p lt; 0.01), significantly higher than the high-dose group (10 U / mL of FSH 20 U / mL LH) maturation rate (26.83%, p lt; 0.05 ). Compared different maturity incubation time of oocyte maturation rate, to mature the oocytes were cultured, respectively, in mature cultured for 24 h, 30 h, 36 h, 40 h on the part of the oocytes with hyaluronic acid first polar body excretion rate was observed after the enzyme treatment, the rest of the fertilization rate plus sperm observed. Cultured 24 h, 30 h, 36 h, 40 h maturation rate of 21.92%, 46.62%, 52.77%, and 30.71%, respectively, which show that: maturation time of 30,36 h of maturation rate was significantly higher than mature 24 40 h the maturation rate (p lt; 0.05). TCM199 FCS FSH LH E2 sodium pyruvate basic maturation medium were added to 50 ng / mL and 100 ng / mL EGF, 10 ng / ml and 20 ng / mL SCF, 10 ng / mL and 20 ng / mL LIF, EGF, SCF and LIF oocyte maturation rate and then compared. The results showed that: 50 ng / mL and 100 ng / mL EGF can significantly with improved oocyte cells mature rate (the basis of mature culture liquid and add 50 ng / mL and 100 ng / mL EGF group of mature rate were 49.04% , 61.39% and 65.79%, respectively). SCF had no significant effect on oocyte maturation rate (maturation rate were 54.17%, 50.00% and 45.83%, respectively), while the LIF oocyte maturation significantly inhibited (maturation rate of 54.19%, 12.50% and 4.17% ). 5 compares the upstream method, the direct washing and Percoll density gradient centrifugation three sperm processing methods rabbit sperm recovery rate and cleavage rate, the recovery rate of the three methods were 7.8%, 93.9%, 86.9%, cleavage after fertilization rate of 0%, 39.05%, 54.29%, The results showed that: Percoll density gradient centrifugation is an ideal sperm processing methods. 6 compares the conventional in vitro fertilization and microinjection cleavage rate, the results show the conventional in vitro fertilization cleavage rate (63.97%) was significantly higher than the microinjection cleavage rate (25.65%) (p lt; 0.01). Compared TCM199 10% FCS and DMEM 10? S embryos (obtained from the conventional in vitro fertilization - cells) and development of, the results showed that no significant difference (p gt; 0.05) between. 8. TCM199 10? S fertilized eggs based culture medium, were added to a certain concentration of vitamin C (100μmol / L) and β-mercaptoethanol (50μmol / L), and then compare them to the embryos (obtained from conventional IVF 2 - and development of cells), showed that, 100μmol / L of vitamin C and 50μmol/Lβ--mercaptoethanol on embryonic development no significant effect (p gt; 0.05).
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