Signature Program: Reproductive Sciences

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Signature ProgramReproductive Sciences
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Reproductive Sciences Laboratory (RSL)

• Faculty
• Mark Westhusin
• Duane Kraemer
• Charles Long
• Visiting Scientist
• Mundhir Ridha
• Staff
• Taeyoung Shin
• Christy Bormann

• Graduate Students
• Carrie Hanna
• Jie Liu
• Arantza Lassala
• Jessica Epple-Farmer
• Abigail Casillo
• Lara Charles
• Beth Kidd-White
• Suzanne Mengas
• James Williams

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Affiliations

• Interdisciplinary Faculty of Reproductive Biology
  (IFRB)
• 42 Members, 4 Colleges
• 11 CVM, 6 Joint Appointments in CVM
• Center for Genomics and Biotechnology

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Nucleus Transfer

• Collecting and storing nucleus donor cells
• Surgical procedure
• Live animal
• Post mortem
• Culturing cells
• Cryopreservation (freezing) of cells

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The Technology

• Preparing nuclei for transfer
• Regulating cell cycle
• Serum starvation
• Obtaining recipient ova
• In vivo matured
• In vitro matured

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The Technology

• Micromanipulation

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Nucleus Transfer

• Enucleating ova (removing the nucleus)

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The Technology
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The Technology

• Transfer of the cell

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The Technology
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The Technology

• Cell fusion

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Cell Fusion
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Cell Fusion
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The Technology

• Alternative Method
• Intracytoplasmic nucleus injection

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The Technology

• DNA Reprogramming
• Methylation
• Demethylation

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The Technology

• Activation
• Electrical
• Chemical
• Insitol 1,4, 5-triphosphate
• Ionomycin b-dimethylaminopurine

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Examples of Clones

• Second Chance
• 21 year-old Brahman Bull

Photos courtesy of Hill, et al., Biology of
Reproduction 621135 (2000)
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Second Chance (3 years old)
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Examples of Clones

• 86 Squared
• Brucellosis resistant bull
• Tissue frozen for 15 years

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86 Squared (2 years old)
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Examples of Clones
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White-tailed Deer (Dewey)
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Dewey 16 months old
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Examples of Clones

• CC The first cloned cat

Progenitor
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Current Research Projects

• In Utero stem cell trasplantation to correct
  hemophilia A. NIH subcontract through University
  of Nevada, Reno, 680,500. P.I. of Subcontract
  D.C. Kraemer, 2003-2007

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Objective

• To re-establish and extinct strain of hemophilic
  sheep from 6 straws of frozen semen
• Work done at U.N.-Reno (mobile lab)

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Full Spectrum of Reproducive Technologies

• Multiple Ovulation Embryo Transfer (MOET)
• Artificial Insemination (AI)
• In-Vitro Oocyte maturation (IVM)
• In-Vitro Fertilization (IVF)
• Intra Cytoplasmic Sperm Injection (I CSI)
• Nucleus Transfer Cloning (NTC)

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Rehearsal Experiment

• Procedures
• MOET
• MOETNTC
• IVF
• ICSI
• NTC

Transfers 4 4 3 2 2 15
Pregnancies 3 2 0 0 0 5
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First Experiment

• Procedures
• MOET
• IVF
• ICSI
• NTC

Transfers 0 0 17 2 19
Pregnancies 0 0 11 0 11
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Research Projects

• Somatic cell nucleus transfer in the canine.
  Texas Heart Institute. 72,721. P.I. D.C.
  Kraemer, 2004-2005

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Canine Cloning

• 9 bitches
• Huntsville (Labradors and hounds)
• Local (Basset x Dalmatian)

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Procedure

• Oocyte collection and in vitro maturation (IVM)

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Results to Date

• IVM- collected average 64 MI and MII
• Transfer average 36 couplets
• No full term pregnancies so far

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