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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

Biotechnology: Status and Uses

Animal Science 434

John J. Parrish

reproductive biotechnology s
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
In Vitro Production of Embryos
  • Oocyte isolation and maturation
  • Sperm preparation
  • Sperm capacitation
  • Fertilization
  • Embryo Development
in vivo embryo production
In Vivo Embryo Production
  • Normal cycling female
    • horses
  • Superovulated female
    • Cattle
    • Sheep
    • Goats
    • Deer
    • Humans
stimulating follicular development

PGF2a

eCG orFSH

Multiple

Ovulations

Estrus

Estrus

Progesterone

From C.L.

First Follicular

Wave

10-12

Stimulating Follicular Development
stage of embryo at recovery
Stage of Embryo at Recovery

Tight Morula (day 5 - 7)

Early Blastocyst (day 7 - 8)

Blastocyst (day 7 - 9)

embryo sexing
Embryo Sexing
  • Hy Antigen
    • Associated with male cells
  • PCR and Detection of Y and X DNA
slide11

Octopus Springs - Yellowstone National Park

Home of Thermus aquaticus - Taq Polymerase

sperm gender selection
Sperm Gender Selection

Selection of X or Y sperm

percent dna differences between x and y chromosome
PERCENT DNA DIFFERENCES BETWEEN X and Y CHROMOSOME

Human 2.9

Cattle 3.8

Chinchilla 7.5

Turkey 0

X Chromosome has more DNA!!!

slide15

Flow Cytometer Separation of X and Y Sperm

Sperm

Stained

With DNA

Sensitive

Fluorescent

Dye

9

L

A

S

E

R

-

+

+

-

Y

s

o

r

t

X

s

o

r

t

sperm gender selection16
Sperm Gender Selection
  • Flow Cytometry
    • Only method that works!
    • Very few sperm recovered
    • Reduced fertility
    • Expense will limit use
fetal sexing
Fetal Sexing

OR

Ultrasound Evaluation

  • Day 55 - 65
cloning
Cloning

Split morula

cloning by nuclear transfer
Cloning by Nuclear Transfer

Cycles are limited

Only 3 - 4 cycles

nuclear cloning to produce stem cells
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

stem cells grown in culture and allowed to differentiate
Stem Cells Grown in Culture and Allowed to Differentiate

http://www.ansci.wisc.edu/jjp1/as434/powerpoint/fa07/stem_cells.mov

benefits of stem cell research
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
gene transfer using viral transfection
Gene Transfer Using Viral Transfection

Better success but left with potential for viral replication

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