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Biotechnology: Status and Uses

Biotechnology: Status and Uses. Animal Science 434 John J. Parrish. Reproductive Biotechnology's. Artificial insemination In vitro embryo production In vivo embryo production Embryo transfer Gender selection Stem Cells Genetic engineering Cloning. In Vitro Production of Embryos.

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Biotechnology: Status and Uses

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  1. Biotechnology: Status and Uses Animal Science 434 John J. Parrish

  2. Reproductive Biotechnology's • Artificial insemination • In vitro embryo production • In vivo embryo production • Embryo transfer • Gender selection • Stem Cells • Genetic engineering • Cloning

  3. In Vitro Production of Embryos • Oocyte isolation and maturation • Sperm preparation • Sperm capacitation • Fertilization • Embryo Development As a direct result of research conducted by Parrish, Susko-Parrish, First, Leibfried-Rutledge, Critser and Ax, more than 450,000 IVP embryos are produced world wide each year and the procedures are the among the most cited publications in all of reproductive physiology!

  4. In Vivo Embryo Production • Normal cycling female • horses • Superovulated female • Cattle • Sheep • Goats • Deer • Humans

  5. PGF2a eCG orFSH Multiple Ovulations Estrus Estrus Progesterone From C.L. First Follicular Wave 10-12 Stimulating Follicular Development

  6. Non-Surgical Embryo Flush Day 6 or 7

  7. Stage of Embryo at Recovery Tight Morula (day 5 - 7) Early Blastocyst (day 7 - 8) Blastocyst (day 7 - 9)

  8. ET - Scheme

  9. Embryo Sexing • Hy Antigen • Associated with male cells • PCR and Detection of Y and X DNA

  10. Hy-Antigen

  11. Octopus Springs - Yellowstone National Park Home of Thermus aquaticus - Taq Polymerase

  12. PCR Approach to Embryo Sexing Y X F M F F F F F F

  13. Sperm Gender Selection Selection of X or Y sperm

  14. PERCENT DNA DIFFERENCES BETWEEN X and Y CHROMOSOME Human 2.9 Cattle 3.8 Chinchilla 7.5 Turkey 0 X Chromosome has more DNA!!!

  15. Flow Cytometer Separation of X and Y Sperm Sperm Stained With DNA Sensitive Fluorescent Dye 9 0° 0° L A S E R - + + - Y s o r t X s o r t

  16. Sperm Gender Selection • Flow Cytometry • Only method that works! • Very few sperm recovered • Reduced fertility • Expense will limit use

  17. Fetal Sexing OR Ultrasound Evaluation • Day 55 - 65

  18. Cloning Split morula

  19. Cloning by Nuclear Transfer Cycles are limited Only 3 - 4 cycles

  20. Nuclear Cloning to Produce Stem Cells Cells of InterestGrown in Culture Dish Mature Oocyte enucleated and cell of interest fused Embyro?? Develops • Potential Tissue: • Neural • Adipose • Connective • Heart Differentiate Inner Cell Mass Cells isolated and grown in culture

  21. Stem Cells Grown in Culture and Allowed to Differentiate http://www.ansci.wisc.edu/jjp1/as434/powerpoint/fa07/stem_cells.mov

  22. Benefits of Stem Cell Research • Regenerative medicine and tissue engineeringIn vitro systems: drug discovery, toxicology, diagnostic assays and cell culture reagentsBiomedical research: unravel mechanisms of disease and human development

  23. Genetic Manipulation

  24. Transgenic for Growth Hormone

  25. Gene Transfer Using Micro-Injection of Pronuclei Less than 1% efficiency

  26. Gene Transfer Using Viral Transfection Better success but left with potential for viral replication

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