Frozen: The story of Frostie the calf

Frostie calf CROPIan Wilmut is best known for his involvement with the team which cloned Dolly the sheep in 1996. However, his scientific career, which spans more than five decades, includes a variety of groundbreaking achievements and discoveries, which are being revealed as his papers are catalogued as part of the continuing ‘Towards Dolly’ and ‘The Making of Dolly’ projects.

Ian Wilmut initially wished to work as a farmer, but he ‘shuffled sideways into scientific research’*, as he puts it, and ultimately won a scholarship as an undergraduate at Nottingham University to work for two months under Chris Polge at  the Animal Research Station in Cambridge. His role was to help out generally with experiments, but he soon became fascinated by embryos, and returned to work with Polge once he graduated. Wilmut’s PhD, awarded in 1971, was on the freezing of boar semen. Copies of Wilmut’s published papers exist in the archive from 1969 onwards, and a glance through the papers which appeared over the next decade reveal Wilmut’s wide-ranging research on the effects of freezing, thawing and warming on embryos and spermatozoa in mice, sheep and cattle.

In 1973, Wilmut was the first scientist to successfully freeze a calf embryo (using liquid nitrogen), thaw it, and transfer it to a surrogate mother. This process led to the birth of a healthy red and white Hereford-Friesian cross calf, which Wilmut wryly named ‘Frostie’. Wilmut’s findings were published within a few weeks of Frostie’s birth in The Veterinary Record as ‘Experiments on the low-temperature preservation of cow embryos’ (June 30 1973, 686-690), and a copy of the reprint survives in the archives. Wilmut has remarked that this was ‘one of the fastest scientific publications ever’. It also led to some considerable interest from the world’s media, with Wilmut appearing on television and newspapers from as far away as New Zealand seizing upon the story. This media attention was a precursor to the storm which Wilmut, Keith Campbell and team would generate two decades later when Dolly the sheep was born. Wilmut’s discovery of the viability of frozen embryos to produce healthy offspring has since been used across many different species in agriculture and also for the conservation of rare breeds. The first human to be born from a frozen embryo was Zoe Leyland, born in Melbourne in 1984.

Wilmut’s work with Chris Polge equipped him with many of the techniques in reproductive physiology which would instruct his later work on cloning, nuclear transfer, stem cell and regenerative medicine in Edinburgh, where Wilmut moved in 1973. Throughout his career Wilmut has been inspired by the possibilities of advances in reproductive physiology and biotechnology for fertility treatments,practical applications to the farming industry and breakthroughs in treatments or cures for debilitating genetic diseases.

*All quotes taken from The Second Creation: the age of biological control by the scientists who cloned Dolly, I. Wilmut. K. Campbell and C. Tudge (London, 2000).
Clare Button
Project Archivist

Printed Material Highlights from the Roslin Collection

With phase I of the Towards Dolly project ending for me, the Rare Books Cataloguer, I thought to look back over the material I’ve worked with and to highlight a few of my favourites from the three different mediums – offprints, rare books and glass plate slides. One of the most interesting things I found when cataloguing this material was the range of topics of interest – the geneticists collected a wide-range of subjects from specifically dealing with animal genetics to ethnography and  botany that opens this material to a variety of researchers.

In the offprint series the two – out of thousands – that I’d like to feature are:

RoslinOffprintDollyUpdateFrom the Roslin Institute offprints, Harry D. Griffin’s article, Update on Dolly and nuclear transfer, Roslin Institute, Edinburgh: Annual Report from 1 April 97 to 31 March 98, (GB237 Coll-1362/4/1848) which discusses the advances in nuclear transfer technology a year after Dolly, the sheep’s birth.

 

CrewOffprintWJBryanFrom the FAE Crew offprint series, William Jennings Bryan’s closing argument in the Scopes evolution case in Tennessee from 1925. (GB 237 Coll-1496/33 – General Biology 2). Bryan was the prosecuting attorney in the ‘Scopes Monkey Trial’ who argued against the teaching of evolution in schools with Clarence Darrow for the defence.

 

 

Two from the rare book collection:

Roslin_S_10

The beautifully illustrated cover contains an interesting map, text and illustrations of the Chinese Langshan fowl in A. C. Croad’s book, The Langshan fowl: history and characteristics from 1899. (Roslin.S.10)

 

 

 

Roslin_S_50-58_7Another beautifully illustrated cover, is a favourite of mine from the nine volume series, The horse : its treatment in health and disease, with a complete guide to breeding training and management, 1905 (Roslin.S.50). Some of the volumes have pop-up style inserts showing the physiology of hooves and mouths layer-by-layer.

 

 

And two from the glass plate slides collection:

Ostriches Pigs and PumpkinsThis is one of my absolute favourites from the Roslin glass plate slides collection simply for its oddity – ostriches and pigs in a field of pumpkins with farm houses in the background in the early 20th century. (Coll-1434/1177).

 

 

Coffee Ranch in Vera CruzCoffee is a passion of mine and so I was thrilled to find this image of a coffee ranch in Vera Cruz, Mexico in the early 20th century. The image shows a family standing in front of their thatched hut, a man on a horse and two men carrying coffee bean baskets on their back with trees and bushes in the background. (Coll-1434/1103.

 

Each of these are personal favourites, as well as being a representative sample of the diversity of the collection. I hope you enjoy it as much as I have!

A Sample of the Roslin Institute’s Cloning Research Post-Dolly: 1998 and 2007

Dolly and Bonnie

As mentioned in a previous post, Dolly, the sheep caused a media sensation in 1997 as the first cloned animal using a nuclear transfer process  and so, I thought it would be interesting to highlight several articles that I came across on Dolly and cloning at the Roslin Institute in 1998 and then again in 2006. I wondered what cloning research had developed over the years since Dolly, the sheep’s birth in 1996 and surprisingly, or not, the articles I came across (that evoked Dolly) dealt with the issue of eating cloned animal meat and the ethical debate of cloning humans for medical purposes.

Note:  these four articles are just a sampling of the articles produced by the Roslin geneticists on the  issues, debates and research surrounding Dolly, nuclear transfer, animal and human genetics, cloning purposes (medical, agricultural, genetic conservation, etc..) to illustrate what way being discussed at the time. For more articles on these subject, please consult the Roslin Institute off-prints for 1998 and 2006 at GB237 Coll-1362/4/.

Update on DollyIn the 1998 Roslin off-print bound volumes, I found Harry Griffiths report, ‘Update on Dolly and nuclear transfer’ in the Roslin Institute, Edinburgh: Annual Report April 1, 97-March 31 (GB 237 Coll-1362/4/1848) and Sir Ian Wilmut’s article, ‘Cloning for Medicine’ in Scientific American, December 1998 (GB 237 Coll-1362/4/1897). Griffiths report describes Dolly’s creation by the Roslin geneticists and notes that their breakthrough caused several other groups to ‘take advantage of public interest in cloning to advertise their successes …. Calves cloned from adult animals were reported from Japan and from New Zealand.’ The New Zealand clone was from ‘the last surviving animal of a rare breed’  which highlighted the use of cloning to preserve endangered species. He continues with discussing Intellectual Property issues in relation to Professor Yanagimachi and his colleagues at the University of Hawai’i ‘Honolulu Cloning Technique’ and closes with a couple of paragraphs on human cloning. He notes the UK Human Genetics Advisory Commission and the Human Fertilisation and Embryology Authority’s report ‘Cloning  issues in Reproduction, Science and Medicine’ from 7 December, 1998 which recommends that ‘there should be a continued ban on all ‘reproductive  cloning’ – the cloning of babies – but gives cautious support  to the cloning of human cells for therapeutic purposes.’

IMG_4359Wilmut’s article in Scientific American reports on the how biomedical researchers are developing ways to use genetically modified mammals for medical purposes.  He mentions the sheep, Megan and Morag who were the first mammals cloned from cultured cells. A technique that allows cloned sheep to carry human genes and such animals produce milk that can be processed to create therapeutic human proteins. The sheep, Polly, is a transgenic clone of a Dorset sheep and ‘a gene for a human protein, factor IX, was added to the cell that provided the lamb’s genetic heritage, so Poly has the human gene.

In the 2006 Roslin off-print bound volumes, I found two fascinating articles:– Sir Ian Wilmut’s  ‘Human cells from cloned embryos in research and therapy’ in BMJ Vol. 328, February 2004 and J. Sark, et al.’s  ‘Dolly for dinner? Assessing commercial and regulatory trends in cloned livestock’ in Nature Biotechnology, Vol. 25, No. 1, January 2007.

IMG_4371Sir Ian Wilmut’s article ‘Human cells from cloned embryos in research and therapy’ in BMJ Vol. 328, February 2004 is one of the more contemporary papers in the collection that discusses stem cell technology and human cloning issues. He cites studies of human genetic diseases and how cloned cells ‘will create new opportunities to study genetic disease in which the gene(s) involved has not been identified’, specifically describing work with motor-neurone diseases. Then, Wilmut notes how stem cells could be used in treatments for a variety of degenerative diseases, i.e. cardiovascular disease, spinal cord injury, Parkinson’s disease and Type I diabetes. Finally Wilmut discusses the differences in regulation of nuclear transfer and human cloning in various countries, noting that in the United Kingdom, ‘project to derive cells from cloned embryos may be approved by the regulatory authority for the study of serious diseases. By contrast human reproductive cloning would be illegal.’

Dolly for DinnerThen, in 2007, the article by and J. Sark, et al’s ‘Dolly for dinner? Assessing commercial and regulatory trends in cloned livestock’ in Nature Biotechnology, ‘reviews the state of the art in cloning technologies; emerging food-related commercial products; the current state of regulatory and trading frameworks, particularly in the EU and the United states and the potential for public controversy.’

As you can see by these four examples there are a range of issues and concerns that have been discussed over the years. While advances are made in cloning and genetic modification, there are still ethical debates to be had and more research to be done. In reading over these and other similar articles in the Roslin off-prints, I enjoyed learning about the different uses of transgenic animals.

Dollymania – Seven Days that Shook the World

Dolly the sheep

1997 was quite a significant year for the Roslin Institute with “’Dolly, the sheep, ‘…the first mammal cloned from a cell from an adult animal…generated an amazing amount of interest from the world’s media.” (Griffin, Harry. ‘Dollymania’, University of Edinburgh Journal, XXXVIII: 2, December 1997, GB237 Coll-1362/4/1476). And so, it’s been exciting to find articles in the offprints discussing her and the issues of cloning, biotechnology, ethics – Dr. Grahame Bulfield even wrote a report to Parliament on what this breakthrough means for science!

Harry Griffin, former Assistant Director (Science) at the Roslin Institute in 1997’s article, ‘Dollymania’ (cited above) provides an insider’s point of view of how Dolly was produced and the science and research involved. He writes,

Dolly was produced from cells that had been taken from the udder of a 6-year old Finn Dorset ewe and cultured for several weeks in the laboratory. Individual cells were then fused with unfertilised eggs from which the genetic material had been removed and 29 of these ‘reconstructed’ eggs – each now with a diploid nucleus from the adult animal – were implanted in surrogate Blackface ewes. One gave rise to a live lamb, Dolly, some 148 days later. Other cloned lambs were derived in the same way by nuclear transfer from cells taken from embryonic and foetal tissue.

On Monday, Dolly provided the lead story in most of the papers and Roslin Institute was besieged by reporters and TV crews from all over the world…. Dolly rapidly became the most photographed sheep of all time and was invited to appear on a chat show in the US. Astrologers asked for her date of birth and PPL’s share price rose sharply. President Bill Clinton called on his bioethics’ commission to report on the ethical implications within 90 days and Ian Wilmut was invited to testify to both the UK House of Commons and the US Congress…. Dolly Parton sad she was ‘honoured’ that we have named our progeny after her and that there is no such thing as ‘baaaaaed publicity’. Sadly, we also received a handful of requests to resurrect relatives and loved pets.

IMG_4284In the article, ‘Seven days that shook the world’ by Harry Griffin and Ian Wilmut in New Scientist, 22 March 1997 also describe the reality of the science of cloning in the face of intense media speculation and reportage.

Dr. Grahame Bulfield, former Director of the Roslin Institute, wrote several articles in 1997 on biotechnology, ethics, livestock and cloning. In some articles, he writes generally on the techniques of genetic engineering, genome analysis, and embryo manipulations and provides a biological context of these new technologies. (GB237 Coll-1362/4/1394 – 1400). He discusses Dolly more directly in the article, ‘Dit is pas het begin’ in the Dutch journal Natuur & Techniek, No. 8, 1997 (4/1376) IMG_4266and The Roslin Institute and Cloning an address to the Parliamentary and Scientific Committee in Science in Parliament, Vol. 54, No. 5, September/October 1997 (4/1400). In this particular article he writes specifically about Dolly:

As you know we have been thrown into the middle of public debate recently with a considerable amount of public interest and concern about “Dolly”. IMG_4275Over a period of about five days we had 3,000 telephone calls, 17 TV crews and we basically ground to a halt. We are perfectly aware now of the issues that are raised , and I don’t believe that a scientific organisation like ours can do anything buy try and be proactive in terms of communicating new biotechnology advances to the public and Government and ensuring the issues involved are widely debated.

Dolly, Polly, Molly, Megan and Morag

Dolly and press Murdo MacleodHere on the ‘Towards Dolly’ team we couldn’t let the 05 July go by without celebrating our namesake, who was born on this day in 1996. To most people, Dolly the sheep (1996-2003) needs no introduction. The first mammal to be cloned from adult cells, Dolly was produced at the Roslin Institute, Edinburgh as part of research into producing medicines in the milk of farm animals. The creation of Dolly met with public acclaim and outcry, fuelling the continuing debates surrounding the ethics of cloning. Most people know the basics about Dolly (including, of course, how she acquired her name), but here are a few facts that may surprise:

Dolly’s birth was kept under wraps for seven months

Dolly’s birth was announced to the world in Nature (385, 753-844) on 27 February 1997, when Dolly was already seven months old. (This time delay was so that the research could be properly prepared for presentation.) Although the journal featured ‘Dolly’ on the front cover, the ‘announcement’ was couched in somewhat muted terms: ‘Viable offspring derived from fetal and adult mammalian cells.’ It is only towards the close of the article that the phrase ‘The lamb born after nuclear transfer from a mammary gland cell is, to our knowledge, the first mammal to develop from a cell derived from an adult tissue’ suggests the importance of the event. Even so, few people at Roslin realised what the strength and duration of public interest in Dolly would be.

 Clones pre-Dolly

Of course, there are naturally occurring clones in nature, such as in bacteria. In terms of laboratory cloning, transgenic frogs, mice and cows have been available from the 1980s onwards. The difference with Dolly was that it is so much more difficult to clone from an adult cell. Dolly was the only live lamb to emerge from 277 attempts.

Ever heard of Megan and Morag?

Megan and Morag were identical twin sheep cloned from the same embryo and were the first mammals to have been successfully cloned from differentiated cells. They were born at the Roslin Institute in July 1995. Although Megan and Morag never got the same level of publicity as Dolly was to have, there was a lot more resting on their birth. Crucially, Megan and Morag demonstrated that viable sheep can be produced by nuclear transfer from cells which have been cultured in vitro. The technical breakthrough which produced them made Dolly the sheep possible.

Polly and Molly the sheep

Since Dolly, other sheep have since been cloned from adult cells, as have cats, rabbits, horses, donkeys, pigs, goats and cattle. In 1997, two ewes were born at Roslin which were the first mammals to have been successfully cloned from an adult somatic cell (like Dolly) and to be transgenic at the same time. Scientists used cells into which a new gene had been inserted so that the animals produced a therapeutic protein in their blood. Polly and Molly built on the work that had been done with Dolly to demonstrate the therapeutic potentials of recombitant DNA technology combined with animal cloning.

Read more about Dolly and the work of the Roslin Institute here: http://www.roslin.ed.ac.uk/public-interest/dolly-the-sheep/

Over the next two years, I will be cataloguing the papers of Professor Sir Ian Wilmut (part of the team who cloned Dolly), Professor Grahame Bulfield (former Director of Roslin) and the Roslin Institute itself as part of the second phase of our Wellcome Trust-funded project. I look forward to discovering more about Dolly the sheep as well as the work of the Roslin Institute from its inception in 1993 until the early 21st century.

 Photograph reproduced with kind permission of Murdo Macleod.