Selected rearch technique_ppt

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Presentation of Experimental Results 1 Selected Research Technique Research supervisor Dr. Rangsun Parnpai Associate Professor School of Biotechnology, IAT, SUT Team members Thabtim M5430154 Tayita D5430024 Theesit D5430031 Ashit D5430246 E-mail: akpaul2008@gmail.com

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Objectives To compare the fertilizing ability and developmental potentiality to blastocyst stage in between fresh and vitrified grade A oocytes To compare the maturation rate in between grade A and B oocytes To compare the fertilizing ability and developmental potentiality to blastocyst stage in between grade A and B oocytes

Techniques:

Techniques 3 Oocyte fixation and staining In vitro maturation(IVM) In vitro fertilization(IVF) In vitro culture(IVC) Vitrification ( Cryotop )

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4

Experiment I:

Experiment I Comparisons of maturation potential between Grade A and B oocytes Oocyte collection Grade B Oocyte Grade A Oocyte IVM for 23 h Denude and observe 1 st PB Oocyte fixation and staining for check chromosome Check 1 st PB extrude and stage of oocyte

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6 1 st polar body Experiment I

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7 GV GVBD A I GV MI

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8 M II T I Experiment I

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9 Oocyte No. of oocytes M II (1 st PB) MII (Fixing and staining) Non MII oocytes GV (%) GVBD (%) AI (%) TI (%) MI (%) A 60 53.3 a 80.0 a 0.0 a 0.0 3.3 a 8.7 8.0 B 60 36.7 b 57.0 b 17.0 b 3.3 8.3 b 5.0 10.0 a,b Values within the same column differ statistically (P<0.01) Table 1 . Comparisons of maturation potential between Grade A and B oocytes. Experiment I

Conclusions:

Conclusions The maturation rate of grade A oocytes is significantly higher than grade B. Fixing and staining is a good technique for identification of stages oocytes . 10 Experiment I

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11 Experiment II

Experiment II:

Experiment II Comparisons of fertilizing ability and developmental potential to blastocyst stage between Grade A and B oocytes Oocyte collection Grade B Oocyte Grade A Oocyte IVM for 23 h IVF IVC for 8 days Sperm preparation % Blastocyst % 8-cell % Morula % Clevage

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13 2-4 Cell stage 8-Cell stage Morula stage Blastocyst stage 8 Cell stage

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14 Oocytes grade No. of oocytes Cleavage rate (%) Embryo 2-4C (%) 8C (%) Morula (%) Bl (%) A 60 81.7 a 33.3 a 45.0 a 36.7 a 26.7 a B 60 33.3 b 16.7 b 16.6 b 11.7 b 8.3 b a,b Values within the same column differ statistically (P<0.01) Table 2. Comparisons of fertilizing ability and developmental potential to blastocyst stage in between Grade A and B oocytes. Experiment II

Conclusion:

Conclusion The Grade A oocyte has significantly higher fertilizing ability and developmental potentiality to blastocyst stage compared with grade B. 15 Experiment II

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Comparing fertility and blastocyst rate in fresh and vitrified grade A oocyte Oocyte collection IVM for 22 h IVF IVC for 8 days Vitrification ( cryotop ) Thawing oocyte and culture in IVM for 1 h FDA staining and select only alive oocyte for IVF Fresh oocyte % 8-cell % Morula % Blastocyst % Clevage Experiment III

FDA staining:

FDA staining 18 Fresh Vitrified Experiment III

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19 Group No. of oocytes Survival rate (%) Cleavage rate (%) Embryo development 2-4 C (%) 8 C (%) Morula (%) Bl (%) Fresh 60 92.4 78.4 a 35.0 43.3 a 38.3 a 28.3 a Vitrified 60 87.3 68.3 b 33.3 35.0 b 30.0 b 21.7 b a,b Values within the same column differ statistically (P<0.05) Table 3. Comparison of fertilizing ability and developmental potential to blastocyst stage between fresh and vitrified grade A oocytes. Experiment III

Conclusion:

20 The fresh oocyte has significantly higher fertilizing ability and developmental potentiality to blastocyst stage compared with vitrified. Conclusion Experiment III

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21 Thank You Everybody

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