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Biotechnology: Status and Utilizations

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Biotechnology: Status and Utilizations

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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

  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