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

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Presentation on theme: "Biotechnology: Status and Uses Animal Science 434 John J. Parrish Animal Science 434 John J. Parrish."— Presentation transcript:

1 Biotechnology: Status and Uses Animal Science 434 John J. Parrish 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 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 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 Normal cycling female –horses Superovulated female –Cattle –Sheep –Goats –Deer –Humans

5 Stimulating Follicular Development Estrus eCG or FSH Progesterone From C.L. Progesterone From C.L. First Follicular Wave First Follicular Wave 10-12 Multiple Ovulations Multiple Ovulations PGF 2 

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 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 X Y 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 Human 2.9 Cattle 3.8 Chinchilla 7.5 Turkey 0 X Chromosome has more DNA!!!

15 + + - - + - L L A A S S E E R R 90° X X s s o o r r t t Y Y s s o o r r t t Sperm Stained With DNA Sensitive Fluorescent Dye Sperm Stained With DNA Sensitive Fluorescent Dye Flow Cytometer Separation of X and Y Sperm

16 Sperm Gender Selection Flow Cytometry –Only method that works! –Very few sperm recovered –Reduced fertility –Expense will limit use 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 Interest Grown in Culture Dish Cells of Interest Grown in Culture Dish Mature Oocyte enucleated and cell of interest fused Embyro?? Develops Embyro?? Develops Inner Cell Mass Cells isolated and grown in culture Differentiate Potential Tissue: Neural Adipose Connective Heart Potential Tissue: Neural Adipose Connective Heart

21 Stem Cells Grown in Culture and Allowed to Differentiate

22 Benefits of Stem Cell Research 1.Regenerative medicine and tissue engineering In vitro systems: drug discovery, toxicology, diagnostic assays and cell culture reagents Biomedical 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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