Snuppy in the wind. Next: Family Photo: Snuppy (Right) sits next to the adult male brindle Afghan hound (Left) from which he was cloned. Snuppy's surrogate mother actually was a yellow Labrador Retriever (See main news page).

News: Snuppy Rewards Dogged Approach: Canine Eggs Are Tough To Crack, But Clone Finally Survives"
by
Emma Marris

Slow Clones

Since Dolly the cloned sheep made her appearance nearly a decade ago, the field has not advanced as much as was expected at first. If you come here, if you meet him, you may fall in love with him. Cloning animals has proven difficult and every species presents its own problems. Even when everything seems to be going right, the majority of cloned embryos fail because their genes are expressed in abnormal ways. For dogs, the main challenge lies in harvesting the eggs. Canine eggs leave the ovary at a very early stage of development and then mature as they travel towards the uterus in the oviducts. Harvesting the eggs at the point of ovulation and trying to mature them in a test tube failed. So the research team had to wait and remove the eggs by flushing them out of the oviducts using a custom-made solution. The nucleus of each of these egg was removed and replaced with a nucleus from an ear cell. Successfully fused cells were then implanted in female [genetically-unrelated surrogate] dogs.

Pup Talk

The painstaking work was completed by the lab of Woo Suk Hwang, the South Korean researcher who is famous for creating a cloned human embryo and deriving stem cells. The lab's dog work is reported in this week's Nature. It took a team of about 15 people two-and-a-half years to produce the dog, whose name 'Snuppy' is short for Seoul National University puPPY.

"He is very cute. If you come here, if you meet him, you may fall in love with him," says Hwang, who adds that while the puppy looks exactly like the somatic cell donor, it's still unclear whether their personalities are similar. The donor dog belongs to a Professor of Internal Veterinary Medicine at the university, but the puppy "belongs to all human beings, not to myself or the somatic-cell-donor owner," says Hwang.

Barking Up the Wrong Tree?

Hwang says the point of bringing Snuppy into the world is to pave the way for a line of dogs that could model certain human illnesses. But cloning a kennel of diseased canines is a long way off, according to Mark Westhusin, a Reproductive Biologist at Texas A&M University, College Station, who is famous for cloning a cat. Westhusin and his team spent three years trying to clone a dog before they threw in the towel. He says the Korean sucess, while wonderful, may not have been worth it. "It's a logistical nightmare to work with this species. We've known and predicted for years that it was feasible. It's just, how much time and money and effort do you want to devote to the project?" Westhusin would like to see work done that would make the process easier, such as the development of a hormone treatment to induce canine ovulation, or a method for maturing eggs in a test tube. The current work impresses him more for its single-minded tenacity than for its novelty. "They're an outstanding lab and I'm happy for them, but all this says is that you can clone a dog, which we've basically known for some time."

"Scientists Clone Man's Best Friend"

Wednesday, August 3, 2005; Denver, CO ( AP and CNN) -- Scientists for the first time have cloned a dog. But don't count on a better world populated by identical, well-behaved canines just yet. That's because the dog duplicated by South Korea's cloning pioneer, Woo Suk Hwang, is an Afghan Hound, a resplendent supermodel in a world of mutts, but ranked by dog trainers as the least companionable and most indifferent among the hundreds of canine breeds.

[ Editor's Note: I have personally raised and bred (Black-Masked Red) Afghan Hounds for more than 15 years and the above statement is patently false. I once worked in a generic dog Kennel for six months with all sorts of canine and cat breeds, so I can make this statement with some authority. -- Steve Coles]

The experiment extends the remarkable string of laboratory successes by Hwang, but also re-ignites a fierce ethical and scientific debate about the rapidly advancing technology. Last year, Hwang's team created the world's first cloned human embryos. In May, they created the first embryonic stem cells that genetically match injured or sick patients.

Researchers nicknamed their cloned pal Snuppy, which is shorthand for "Seoul National University Puppy." One of the dog's co-creators, Prof. Gerald Schatten of the University of Pittsburgh School of Medicine, describes their creation, now 14 weeks old, as "a frisky, healthy, normal, rambunctious puppy."

Researchers congratulated the Korean team on improving techniques that might someday be medically useful. Others, including the cloner of Dolly the sheep, renewed their demand for a worldwide ban on human reproductive cloning. "Successful cloning of an increasing number of species confirms the general impression that it would be possible to clone any mammalian species, including humans," said Dr. Ian Wilmut, a reproductive biologist at the University of Edinburgh, who produced Dolly nearly a decade ago.

Since then, researchers have cloned cats, goats, cows, mice, pigs, rabbits, horses, deer, mules, and a gaur, a large wild ox of Southeast Asia. Uncertainties about the health and lifespan of cloned animals persist; Dolly died prematurely in 2003 after developing cancer and arthritis.

"The ability to use the underlying technology in developing research models and eventually therapies is incredibly promising," said Robert Schenken, President of the American Society for Reproductive Medicine. "However, the paper also points out that in dogs as in most species, cloning for reproductive purposes is unsafe." The experiment's outcome only seems to buoy the commercial pet-cloning industry, which has charged up to $50,000 (euro41,000) per animal. The first cloned-to-order pet sold in the United States was a 9-week-old kitten produced by the biotech firm, Genetic Savings & Clone, Inc. of Sausalito, CA.

Company officials said they expect to commercially clone a dog within a year using eggs collected from spaying procedures at veterinary clinics. The South Korean researchers can surgically remove eggs from research animals with fewer regulations than in the United States. "This justifies our investment in the field," said spokesman Ben Carlson. "We've long suspected that if anyone beat us to this milestone, it would be Dr. Hwang's team -- due partly to their scientific prowess, and partly to the greater availability of canine surrogates and ova in South Korea." But the dog cloning team tried to distance its work from commercial cloning. "This is to advance stem-cell science and medicine, not to make dogs by this unnatural method," Schatten said.

On scientific terms, the experiment's success was mixed. More than 1,000 cloned embryos were implanted into surrogate mothers and just three pregnancies resulted.. That's a cloning efficiency rate lower than experiments with cloned cats and horses. Details appear in Thursday's issue of the journal Nature. Like Dolly and other predecessors, Snuppy was created using a method called Somatic Cell Nuclear Transfer, or SCNT.

Scientists transfer genetic material from the nucleus of a donor adult cell to an egg whose nucleus -- with its genetic material -- has been removed. The reconstructed egg holding the DNA from the donor cell is treated with chemicals or electric current to stimulate cell division. Once the cloned embryo reaches a suitable stage, it is transferred to the uterus of a surrogate where it continues to develop until birth. Why have canines been so problematic compared with other mammals? Apparently, because their eggs are released from the ovary earlier than in other mammals. This time, the researchers waited and collected more mature unfertilized eggs from the donors' fallopian tubes.

They used DNA from skin cells taken from the ear of a 3-year-old male Afghan hound to replace the nucleus of the eggs. Of the three pregnancies that resulted, there was one miscarried fetus and one puppy that died of pneumonia 22 days after birth. That left Snuppy as the sole survivor. He was delivered by C-section from his surrogate mother, a yellow Labrador Retriever. Researchers determined that both of the puppies that initially survived were genetically identical to the donor dog.

Schatten said the Afghan hound's genetic profile is relatively pure and easy to distinguish compared with dogs having more muddled backgrounds. But dog experts said the researchers' choice of breed choice was disquieting. "The Afghan hound is not a particularly intelligent dog, but it is beautiful," said psychologist Stanley Coren, author of the best-selling manual "The Intelligence of Dogs." He ranked the Afghan hound last among 119 breeds in temperament and trainability. "Many people who opt for the cloning technique are more interested in fashionable looks," he said. "Whenever we breed dogs for looks and ignore behavior, we have suffered."

[ Editor's Note: This last comment is really ludicrous. I can generate contrary data for anybody who cares to know more about this breed. It is true that Afghans are less eager to please humans than other breeds, but they are extremely clever, and can often outwit you in the same way that cats or monkeys can, while most dog breeds don't even bother to try.]

"HEALTH: Korean Scientists Produce World's First Cloned Dog"
by
Nicholas Zamiska and Antonio Regalado, Staff Reporters for The Wall Street Journal

August 3, 2005; Seoul, SOUTH KOREA -- Move over Dolly. They've cloned man's best friend. South Korean scientists, who have been blazing ahead in controversial areas of embryo research, have cleared another hurdle, producing the world's first cloned dog -- an Afghan Hound. The advance won't answer the prayers of dog owners determined to replicate Fido or Fluffy because the team of researchers has said that it is focusing on trying to cure people, not clone pets. What's more, their procedure wouldn't work on a large scale. Still, the technical leap could help jump-start a pet-cloning industry that is still in its infancy. And the breakthrough is certain to further fuel an already heated debate about the ethics of cloning -- human or otherwise.

Snuppy, is the first cloned dog, at age 67 days, with the three-year-old Afghan hound he was cloned from. The unveiling Wednesday of 14-week-old Snuppy is the latest success for Woo Suk Hwang and his team at Seoul National University, which have been vaulted to international prominence since a report last year that they had successfully cloned a human embryo and extracted its stem cells. Snuppy is a blend of the acronym for the university, SNU, and the word "puppy." Details are reported in an article in Thursday's issue of the journal Nature.

The lab's success at creating the first cloned canine is likely to reinforce South Korea's leadership in the field and stoke worries by American scientists that ethical and regulatory concerns may be hampering progress in the U.S. While some of the techniques Dr. Hwang and his team use on human cells, and have now been applied to dogs, remain legal in the U.S., the Administration of President Bush has sought to have them outlawed.

The South Korean team's three-year effort to clone a dog was motivated by an interest in developing new models of human diseases that could be used for testing drugs or other treatments, says Dr. Hwang, who himself owns six dogs, including a Siberian Husky and a Doberman. "There's no hope, no answer, no technology, if we do nothing," he said. "This work is from bench to bedside."

[See below for a brief Review of Major Accomplishments in Cloning since the birth of Dolly, the sheep.]

Ever since scientists in Scotland cloned a sheep named Dolly, in 1996, researchers have pressed forward with attempts to copy other mammals, succeeding with rats, pigs, cows, and horses, among others. But copying dogs has proved far more difficult because their eggs, unlike those of most mammals, mature outside of a dog's ovaries, making it difficult to predict when they have become mature enough to harvest. Copying dogs also opens the door for mass pet cloning in a way that replicating sheep never could.

In the U.S., commercial pet-cloning efforts are led by Genetic Savings & Clone, Inc. , a Sausalito, CA biotechnology company funded by education and health entrepreneur Dr. John Sperling, with its mission the cloning of Missy, Dr. Sperling's dog who died several years ago. "We suspected that if anyone beat us, it would be the Koreans," a company spokesman, Ben Carlson, said Wednesday. He says Genetic Savings & Clone will produce a cloned dog by the end of this year. The company already provides a cat-cloning service at a cost of $32,000 to pet owners, and says it received five orders in 2004 and "less than a few dozen" this year. Mr. Carlson says the cost of cloning a cat has put off many potential customers but that his company's market research shows people would be more willing to pay for a dog. The company hasn't set a price for dog cloning. Already, many have sent samples of their dog's DNA to be frozen and stored at the company's laboratory until the price of the procedure comes down.

The idea of pet cloning has been met with criticism. Animal-rights advocates have tried to introduce legislation in California and other states in the U.S. to ban the practice, calling it cruel -- to both the clones that can suffer from defects and their surrogate mothers, which must have embryos implanted in their wombs. Some scientists say consumers may be misled into thinking they will receive copies of their pets. "To promise people they are going to get Fluffy back is just not right," says Robert Lanza, Vice President of Medical and Scientific Development at ACT, a Worcester, MA company that has cloned cows. Clones can now be created from the DNA of an existing animal or cells that have been stored. But cloning doesn't produce an exact replica: Physical and behavioral traits can be affected by environmental factors, including how the animal develops in the womb.

To achieve their goal, the Korean scientists implanted more than 1,000 cloned embryos into 123 surrogate mother dogs. That produced only three pregnancies, from which one puppy was still-born and a second died shortly after birth. None of the surrogate mothers died, the scientists said.

The researchers say Snuppy has no obvious physical abnormalities, as have some other cloned animals. In his first public appearance Wednesday, he appeared quite frisky, inspecting the enormous crowd of photographers that had come to take his picture. His surrogate mother, a yellow Labrador, as well as his genetic mother, a stately Afghan Hound with a leisurely mane, also made an appearance. To clone Snuppy, scientists extracted the genetic material from a skin cell from the Afghan hound and implanted into an egg harvested from one of the surrogate mothers. They reconstructed the egg and stimulated it to begin dividing, and then implanted it in the uterus of the yellow Lab. Dr. Hwang, 52 years old, started his genetic research at SNU in 1986 with no laboratory, and, soon after, a $5,000 government research grant. Now, he has a team of more than 60 researchers and annual government funding of just under $1 million. His path-breaking research has made him something of a celebrity here -- nearly 200 reporters attended his news conference, and he is sometimes stopped on the street for autographs. Returning from Shanghai on a recent flight, he stepped off the plane in Seoul and saw hundreds of passengers in the terminal start applauding. His laboratory has evolved into a small factory aimed at producing disease-resistant cows and pigs that may someday provide humans with spare organs. On a recent day, 14 people in blue suits and hairnets worked in near silence at a table piled with cow and pig organs collected from slaughterhouses each morning. Picking up walnut-size cow ovaries and peanut-size pig ovaries, they stick needles into them and extract the fluid that contains the unfertilized eggs.

In addition to cows, pigs, and humans, Dr. Hwang has also been working on other species, including foxes, wolves, tigers, and monkeys. More are on the way. During an interview Wednesday in his cramped office, Dr. Hwang sat beneath a giant green filing folder with the word " mammoth" written on it. So far, the team hasn't been able to salvage healthy enough tissue from the long-extinct animal to make a viable clone.

Write to: Nicholas Zamiska at nichlas.zamiska or Antonio Regalado at antonio.regalado@wsj.com.

From Dolly to Today

August 3, 2005; Snuppy, the first cloned dog, is the latest in a series of cloning successes by researchers around the world. Here is a chronological review of the high points:

February 1997 -- Scotland's Roslin Institute announces its researchers have cloned a sheep from cells taken from an adult ewe. The sheep, called Dolly, was the first animal cloned in such a process; previous cloning had involved creating an animal from undeveloped fetal cells in a surrogate mother.

August 1997 -- Privately held cattle-breeding company ABS Global, of Wisconsin, announces it has cloned a calf from stem-cells removed from a cow fetus.

July 1998 -- Researchers at the University of Hawaii say they've successfully cloned dozens of mice.

August 2000 -- Boston biotech-firm BioTransplant, Inc. and Mass General Hospital announce they have cloned a line of miniature pigs, with the intent of creating organs for transplant into humans.

February 2002 -- People familiar with a Texas A&M University research program known as CopyCat say the scientists successfully cloned a domestic cat in late 2001. The cat, whose birth was later confirmed, was named "cc:".

March 2002 -- French scientists announce the first clones of rabbits from adult cells, just in time for Easter.

May 2003 -- Scientists in Idaho, funded by a local grass and seed magnate, announce they have cloned a mule, a sterile hybrid normally produced by breeding a donkey and a mare. The animal, named Idaho Gem, narrowly nosed out competing efforts by teams in Texas and Italy to become the first cloned equine, an accomplishment much sought after by horse experts.

February 2004 -- Researchers at Seoul National University's veterinary college clone human embryos and extract their stem cells. The ground-breaking development is hailed as a watershed in the push to develop custom-tailored transplant organs.

August 2005 -- South Korean scientists at the same university where the human embroyos were cloned say they've cloned the first dog, a brindle Afghan Hound named Snuppy.

[Editor's Note: All of the above material on animal cloning is in the News Section of our website and in a PowerPoint slide in the Resources Section that I have maintained up-to- date for the last five years along with each new breaking-news development. -- Steve Coles]

"Korean Scientists Clone World's First Dog"
by
Karen Kaplan, LA Times Staff Writer

Researchers in South Korea have produced a black-and-tan puppy named Snuppy that was cloned from an adult Afghan Hound, a long-sought feat anticipated by scientists and dog owners alike. The researchers emphasized that their aim is to improve both canine and human health, not to reproduce beloved pets. "Our goal with cloning researchers is to find cures for debilitating diseases and illnesses," said Woo Suk Hwang, one of the lead researchers from Seoul National University. "With the promise of using a homogenous population of cloned dogs, maladies such as hypertension, diabetes, breast cancer, or genetic disorders like congenital cardiac defect can be studied more efficiently."

But companies that plan to offer dog-cloning services were quick to herald the achievement, reported today by the journal Nature. "This validates one of the premises of our business," said Ben Carlson, a spokesman for Genetic Savings & Clone, Inc. The Sausalito, CA company is storing DNA samples from several hundred dogs in anticipation of producing clones for customers as early as next year.

Pet cloning became a reality in 2002, when scientists at Texas A&M University produced a cloned kitten named Carbon Copy. Genetic Savings & Clone, which funded the research, now charges $32,000 for the service. Though the public has been warming to the idea of pet cloning, sentiment against it remains strong. A Gallup Poll conducted in May found that 61 percent of Americans consider animal cloning morally unacceptable, compared with 35 percent who are comfortable with the idea.

Scientists have been trying to clone dogs since shortly after the birth of a sheep, Dolly, in 1996. The technique involves harvesting unfertilized eggs from females, removing the genetic material and replacing it with DNA from an adult donor, usually taken from a skin cell. The manipulated embryo is then implanted into a surrogate mother. But even as teams around the world produced cloned mice, rabbits, pigs, cows, and even a horse, dogs remained elusive. "Probably most important is the difficulty in obtaining good quality eggs," said Duane Kraemer, a Professor of Veterinary Physiology at Texas A&M University who helped produce Carbon Copy. Female dogs come into heat [estrus] only once or twice a year, providing researchers with relatively few opportunities to extract eggs or implant cloned embryos into surrogate mothers. Making matters worse, eggs harvested from canine ovaries are not mature enough to be coaxed into pregnancy. Research teams have been testing a variety of methods for ripening the eggs in the laboratory, as can be done with eggs from other species, but have yet to hit upon a reliable method.

The South Korean team allowed the eggs to leave the ovaries and mature for a few days before flushing them out of the oviduct, a thin [catheter] tube that delivers eggs to the uterus. Retrieving an egg from the oviduct is much more difficult than collecting it from the ovary, said Betsy Dresser, Senior Vice President for Research for the Audubon Nature Institute in New Orleans, LA, which uses cloning to help preserve endangered species. "Getting those eggs out of the oviduct that were 'in vivo' matured was a very good step," Dresser said.

In addition to using improved techniques, the South Korean team also benefitted from trial and error. The researchers transferred 1,095 cloned embryos into 123 surrogate dogs in order to produce three pregnancies, one of which resulted in a miscarriage. The first puppy, delivered by C-section in late April, was named Snuppy, short for "Seoul National University Puppy." A second puppy died of pneumonia 22 days after its birth. The researchers do not think the illness was related to cloning.

Gerald Schatten, a biomedical researcher at the University of Pittsburgh and a coauthor of the study, said the inefficiency of the process suggests that commercial dog cloning for pet owners is still a ways off. "We transferred 1,095 embryos and got one dog," said Schatten, who leads the Pittsburgh Development Center, a biology research institute. "If I were an investor and someone came in and said we have a technique that works at 0.09 percent, I'd say that doesn't sound like a good investment."

Companies agree that cloning methods will have to become much more reliable before they can begin delivering puppies to customers. "We knew that it could be done," said Philip Damiani, Chief Scientific Officer at Genetic Savings & Clone. "It's just a matter of trying to figure out the optimal protocol to make this efficient."

When they do, people like Peter Lowenstein of Oakland are eager to become customers. Lowenstein has adopted several strays in his 50 years, but none worked into his heart like Sneakers, a 70-pound cross between an Irish Wolfhound, Scottish Deerhound, and Labrador Retriever with wiry charcoal hair and yellow eyes. Sneakers passed away last year while she was under anesthesia for a procedure to investigate a spinal problem. By the time he got home from the veterinarian's office, Lowenstein had made up his mind to bank her DNA. A vet collected tissue from inside her mouth, her abdomen, and some internal organs and sent them to Genetic Savings & Clone, who keeps them in liquid Nitrogen. The company charges $1,395 initially and an annual storage fee of $150. "I was so brokenhearted," said Lowenstein, a Chiropractor. "If she had lived a good life and just died of old age, I don't think I would have done it. But it wasn't her time. I didn't have a chance to say goodbye to her."

[ Comment by Dr. Steven B. Harris, M.D. in response to the question as to how the Hwang Lab managed to beat GS&C after they had been working on the problem for more than five years at great expense and were themselves very close to a cloning a dog of their own (still expected before the end of the year):

Every mammal is different when it comes to cloning, and these differences cause expertise in cloning one species to fail to transfer to others, as well as we'd like. For example, if you're a cow- cloner you apparently have to start all over again for mice or cats. Apparently the South Koreans accomplished this feat by brute force and lots of dog surgery (to harvest all those eggs). The canine is a very difficult and expensive research model in the US, largely due to USDA regulations.

I might add that I personally maintain a dog lab myself, so have some relevant experience. In the US, dogs are probably ten times as expensive per animal to keep as experimentally experimental as are cats (and both are orders of magnitude more expensive than rodents). For dogs vs. cats, it's a matter of cage space, room for exercise, waste cleanup, and many, many other variables. The USDA mandates these rules and sets a much higher standard for keeping experimental animals (rabbits, say) than when these animals are raised for food (cows, say). For companion animals or pets, like dogs and cats, it's far more regulated.

In many Asian countries, they EAT dogs (and even cats). They have no USDA or other governmental agency to monitor research-animal status, so the status of dogs is truly as it appears from the fact that it's also a "food" stock. Thus, they could be caged like chickens, if somebody wanted to do so. Go on from there. In retrospect, it is not surprising then that the South Koreans won the race with GS&C as soon as they did.]