[tt] Success with primate cloning

[Hughes, James J.]

http://news.independent.co.uk/sci_tech/article3152325.ece

Cloning: a giant step

For the first time, scientists have created dozens of cloned embryos
from adult primates. But what are the implications of this technical
breakthrough for the future of mankind?

By Steve Connor, Science Editor

Published: 12 November 2007

A technical breakthrough has enabled scientists to create for the first
time dozens of cloned embryos from adult monkeys, raising the prospect
of the same procedure being used to make cloned human embryos.

Attempts to clone human embryos for research have been dogged by
technical problems and controversies over fraudulent research and
questionable ethics. But the new technique promises to revolutionise the
efficiency by which scientists can turn human eggs into cloned embryos.

It is the first time that scientists have been able to create viable
cloned embryos from an adult primate - in this case a 10-year-old male
rhesus macaque monkey - and they are scheduled to report their findings
later this month.

The scientists will also demonstrate that they have been able to extract
stem cells from some of the cloned embryos and that they have managed to
encourage these embryonic cells to develop in the laboratory into mature
heart cells and brain neurons.

Scientists who know of the research said it was the breakthrough that
they had all been waiting for because, until now, there was a growing
feeling that there might be some insuperable barrier to creating cloned
embryos from adult primates - including humans.

The development will not be welcomed in all quarters. Opponents of
cloning will argue that the new technique of manipulating primate eggs
to improve cloning efficiency will lead to increased attempts at
creating - and destroying - cloned human embryos for research purposes.

Although it is illegal in Britain to place any such cloned embryos into
the womb of a woman, many people also fear that the relative ease of
being able to perform cloning using the skin cells of an adult will
increase the chances of its being applied to produce a cloned baby.
Scientists in South Korea reported in 2004 that they had created the
first cloned human embryo but in 2006 their study was retracted after it
emerged that its main author, Hwang Woo-suk , had committed fraud.

There has only been one other documented example of a human embryonic
clone, but it died after a few days and did not produce stem cells. The
work has so far not been replicated.

The scientists who carried out the latest primate work are believed to
have tried to implant about 100 cloned embryos into the wombs of around
50 surrogate rhesus macaque mothers but have not yet succeeded with the
birth of any cloned offspring.

However, one senior scientist involved in the study said that this may
simply be down to bad luck - it took 277 attempts, for instance, to
create Dolly the sheep, the first clone of an adult mammal.

The work was led by Shoukhrat Mitalipov, a Russian-born scientist at the
Oregon National Primate Research Centre in Beaverton. Dr Mitalipov
helped to pioneer a new way of handling primate eggs during the cloning
process, which involved fusing each egg with a nucleus taken from a skin
cell of an adult primate.

Dr Mitalipov said he was unable to comment on the study until it was
published in the journal Nature. But he told colleagues at a scientific
meeting this year that he had made two batches of stem cells from 20
cloned embryos and tests had shown they were true clones.

Professor Alan Trounson of Monash University in Australia said Dr
Mitalipov's findings represented the long-awaited breakthrough. Despite
many attempts, no one had been able to produce cloned primate embryos
from adult cells, yet this had been done on dozens of other non-primate
species. " This is 'proof of concept' for the primate. It has been
thought by some [to be too] difficult in monkeys - and humans - but
those of us who work [with] animals such as sheep and cattle thought
that success rates would be much like that achieved in these species,"
Professor Trounson said.

"Mitalipov's data confirms this. They have the skills necessary and we
can now move on to consider what might be able to be achieved in
humans."

Professor Don Wolf, who led the laboratory at the Oregon National
Primate Research Centre before his recent retirement, said the new
procedure was based on a microscopic technique that does not use
ultraviolet light and dyes, which appear to damage primate eggs.

"In the early days we tried to use that technique in the monkey and
unbeknownst to us at the time that was basically damaging the egg. So
one of the keys was to remove that step from the process," Dr Wolf said.

"We could now produce cloned blastocysts [embryos] in the monkey at a
reasonable frequency, at least a frequency that would allow us ...to
study the cloned blastocyst ," Professor Wolf said.

The Oregon team, working with a group in China, has so far produced
about 100 cloned embryos that have been transferred into around 50
female macaques, but none has resulted in a full-term pregnancy, he
said.

"It's possible that we're still just having bad luck. We're producing
may be one in 20 or one in 30 cloned blastocysts that are 'normal' and
capable of producing a pregnancy and we just haven't got them into the
animal recipient at the right time to allow implantation and pregnancy
to occur," Professor Wolf said.

"The focus now is going to be on therapeutic cloning and using the
non-human primate as a paradigm for therapeutic cloning for what you
might be able to do clinically," he said.

"We're the first to do it, although it's a tainted subject because of
the fraudulent research that came out of South Korea. One can never be
sure but there may be some validity to what the South Koreans did. But
this would now be the first documented therapeutic cloning in a
primate," he added.

A brief history of cloning

The monkey-cloning technique is the same basic procedure that resulted
in Dolly the sheep. The nucleus of a healthy, unfertilised egg is
removed and another nucleus from the mature skin cell of an adult animal
is placed inside the egg. With careful timing and the use of electrical
pulses, an embryo can be created which is a genetic clone of the skin
tissue donor. It is possible to implant embryos created in this way into
the womb to produce cloned animals. This so-called 'reproductive
cloning' of humans is illegal in Britain and many other countries.
However it has been applied to a range of animal species, including:

* Cow: Many domestic cattle have been successfully cloned. First attempt
to clone an endangered species was Noah, a rare gaur ox, which was
cloned in the US in 2001 but died 48 hours after birth

* Mouse: Cumulina was a common brown house mouse, cloned from adult
cells at the University of Hawaii in 1997. She survived to adulthood and
produced two litters, before dying in May 2000

* Horse: Called Prometea, the first cloned horse, born in Italy in May
2003

* Cat: A kitten called CopyCat was born in 2002 in Texas, and gave birth
to three kittens by a natural father in September 2006

* Dog: Snuppy, born in South Korea. Doubts about its authenticity were
dispelled by DNA tests. The group has also cloned two wolf cubs, called
Snuwolf and Snuwolffy using the same procedure. Cloned Afghan hounds
named Bona, Peace and Hope have also been born