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Lecture 6

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BIOL 1010
James Cheetham

Animal Biotechnology Tuesday, October 19, 2010 - When you search for embryonic stem cell case (spinal cord injury), you find a whole lot of business journals- huge interest among investment community - Clinical trial - Subsequent trials will tell whether it can really work - Promises of animal biotechnology and stem cells o Conserve environment- get animals to live on small acreage o Advance human health- we use a lot of animal models - Timeline o Artificial insemination (cattle, horses)- 1950s  Frozen bull semen and embryos o Embryo transfer- early 1980s  Fertilization- in vitro (in glass, artificial)  * in vivo- in living organism  Egg and sperm, mixed outside body o Embryo splitting and cloning- early 1990s  Embryo formed- can split into two bunches of cells or more  This is how you get fraternal twins o Genetic Preservation  This is when you freeze tissue and germ cells from highly-prized animal o Somatic Cell Nuclear Transfer  Dolly was made using this  Big one- cloning o Transgenic technology  Being researched now  Transferring genes among species- make lots of copies - Dan Gardner- columnist for Ottawa Citizen- predictions usually wrong o McGrath in 1987 was wrong - Somatic= body - Nuclear=nucleus - Transfer - Ian Wilmut- runs institute in Scotland where they cloned sheep o Possibility of cloning humans- evil - Cloning is essentially copying o Identical twins are clones o Plant cells all have germline potential- can break off part and grow another copy from it, while humans just have sperm and eggs- our skin cells and such can’t make another human o Stem cells can - Yeast in same way - Plant clones o Seeds- not cloned- have genes mixed from male and female plant o Cutting is cloning plant o Seedless plants reproduced as clones- bad- less genetic diversity- if disease comes along, can’t survive - Animal clones o Slime mould buds off o Parthenogenesis for some vertebrates- create full-blown organism with just egg (no sperm)- happens for some lizards and small animals naturally, not naturally in mammals o Japanese group did it for mice- induce egg to double chromosome number o Also done for monkeys o Done with human egg cells in culture - Natural human clones o Every cell in body has same genes o Environment also controls developments- fingerprints controlled environmentally o Will not experience same environment, even in uterus - Making clone is simple in theory o Multicellular embryo- break it up and each cell can form whole organism- since 50s o Technically difficult- hundreds of tries for mammals o Can’t just scrape cheek cell and grow human - Barriers o Differentiated cells o Hayflick limit- limit on how many times cell can divide o Higher for baby than old person o Cells differentiate- become more specialized o Scotland- used fully differentiated adult cells (mammary glad) for Dolly- miraculously could regrow entire sheep from it - Potency- stem cell term- how many different cell types this cell can become - Totipotent- can become any type of cell - Nucleus needs particular environment to divide - Early embryo- blastomere - Can reproduce placenta, hair, liver, etc. - Science- cloning mammal: breakthrough of the year o Udder cells- grow in dish- starve and reset- extract DNA o Remove nucleus from black-faced sheep- introduce DNA- embryo o Transplant embryo into black-faced sheep- normal gestation o 277 clones to get Dolly- inefficient o Micromanipulators difficult to use- fine motor skills o Fully differentiated somatic cell from adult- can clone - Method o 1) Quiescent- quiet, not growing well  Starve cells o 2) Collect nucleus o 3) Prepare- glass tube into it, suck nucleus out (in old days, used suction) o Etc. - Took mammary gland cells out- bottle full of nutrients (cell growth medium)- culture (growing cells in culture) - Starve- harvest quiescent cells - Hold cell in place with pipette (suction)- another pipette sucks nucleus out - Egg donor- extract egg- pipette to hold in place- another pipette sucks that nucleus out o Enucleated egg o Nucleus from other cell- shoot in - Electrodes to initiate division/development - Zygote gets bigger and divides into two, four, etc. - Up to Embryo stage - Surrogate mother- morula- block of cells - Oocyte- egg - Study of development- egg to baby- genetically identical organisms help explore environment - Propagation of valuable organisms- copy good animals - Control over reproduction- specific baby qualities - Production of recombinant organisms- goats that make spider silk- difficult to make- copy - Production of engineered organs- organs to be transplanted into humans (humanized) - Cloned animals in food supply- midlevel controversy o FDA took the lead o Funny because they voted to delay FDA’s decision - Protect endangered species - Extinct animal- wooly mammoth o More mammoth ivory selling than elephant ivory- popular (glaciers in Siberia melting- finding many of these) o Russians in time of Tsar (starving) would thaw out mammoths and eat o Suggested candidate for cloning- found in permafrost in Russia o Ice crystals in tissue can rupture, destroy DNA o Australian group working on cloning mammoth o They have lived here (where there’s snow), not in Africa - Genotype of individual- can predict what they’ll come out like - Human factor IX- blood clotting factor that sheep produces - Normally purified from blood- small amount- can get contaminated - Biopharming- using animals to produce biopharmaceuticals o Changes our relationship to animals - Xenotransplantation- xeno (other)- across different species - Hwang faked data- article retracted from Nature- negative light - Genetic Savings & Clone, Inc. o Went out of business o California\ o Commercial pet banking and cloning, 2006 o First cloned cat- Sisi o Store pet’s DNA if you want to clone in future o $5,000 to clone animal o Cloning cats isn’t easy- colouration determined during development especially in female cats o Contributed a lot to research on dog cloning in Texas o Making money off of it - GE pets o Glow fish- can buy from pet store in US o Fluorescent proteins- originally made to detect environmental toxins (glow) o But people liked glowing fish- sold them o Could use as bait when fishing- got loose in environment o Bunny- Alba- transgenic art o Expect to see more of this in future o Human bodies preserved in plastic- German art exhibit- scary- interaction between art and science - Genes influence behavior and cognition, but identical isn’t totally identical - Separate experiences and personalities - Could have temporal separation- article about woman who gave birth to sister o Mother donates eggs before chemo- frozen o Daugher infertile o Mother gives eggs to daughter o Daughter gi
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