“I’ve Cloned Human Embryos”

„I cloned a human embryo“

Cloning humans for the sake of procreation: This taboo has first been broken on March 14, 2003 – by a scientist, who has now broken the silence in an interview with P.M. magazine. Our report shows, that laws will not stop biotechnological progress.

By Sascha Karberg

In the morning of March 14th, 2003, Karl Oskar Illmensee sets off to clone a human being. The early riser who strolls down the lane in Lexington, Kentucky, lost in thoughts, is a well-known embryologists worldwide. He is heading towards a laboratory in a secret place. Around seven a.m. Illmensee – as he is going to tell me four years later – will prepare an experiment that could change the world.

Many researchers have repeatedly stated that they were capable of cloning humans: An aged physician from Chicago, the controversial Italian reproduction expert Severino Antinori, and the Raelian sect that believes in UFOs. They all showed to be nothing but impostors, says the stemcell researcher Hans Schöler of Max-Planck-Institute for Molecular Biomedicine in Münster. Illmensee, however, was "a completely different breed". For a long time the 68 year-old scientist has been famed by his colleagues for possessing "an exeptional hand". He is said to be the first man to successfully clone a mammal – 16 years before Dolly the sheep came into existence. Then, he was accused of forgery and fell in disgrace. Ever since, the now retired researcher has kept his mouth shut. Even his trials to reproductively clone humans he never mentioned in public.

But now the two of us are sitting at a stony beach of Patras in Greece – and the man in the plastic chair opposite me finally breaks his silence. In his back the turqoise sea of the Peleponnes ripples ashore. Every now and then he sips his diet-coke and talks: about cloning and his life – two things inextricably linked to each other. Only the slight movements of his legs and hands bespeak of his nervousness. He talks about triumph and defeat, about pride and fall. And about his personal fall of man on March 14th, 2003 – back in that secret lab of the reproduction physicist Panayiotis Zavos from Lexington, the initiator and supporter of Illmensees experiments.

Illmensee is present when oocytes from an American woman are extracted for the cloning process. With his own hands he places the cells in the saline solution, which he has prepared in the morning, and in which the nuclear transfer, the process of cloning itself, is to take place. He settles in front of a microscope and uses a delicate glass pipette to draw the genome from the first oocyte. He waits for an hour and then places a somatic cell of the woman between hull and membrane of the oocyte. Then he sends electric impulses into the solution twice, each taking only milliseconds. The membranes of both cells melt into each other, the nucleus of the somatic cell with all its genetic information moves into the oocyte. Chemical substances are then added in order to inspire the newly constructed cell to divide. Illmensee carefully places the petri dish into the incubator. Within only a few days, he hopes, a human embryo will evolve from the manipulated cell. The first cloned human. Illmensee hardly sleeps that night. Will the experiment work out? Will a cloned embryo develop? Will the woman get pregnant after the artificially constructed embryo will have been placed into her uterus? And what if: How will the public react to such a sensation? The first cloned baby would make him legend as a researcher. Or at least exonerate him from the reproach of forgery that cost him twenty years of his scientific career.

The next day, he checks on the petri dish and jots down in his lab diary that big dots have become visible in two of ten egg cells. In a third cell, the nucleus is not quite situated in the right spot. 40 hours later, Illmensee notices a five-cell-embryo. Another day later, 64 hours have passed by now, the cloned embryo consists of eight cells. Illmensee proudly writes down: "It is the first!" But the

uterus of the American woman, that attempted to let clone herself, proves to be unsuitable for embryonic transfer – the embryo is therefore deep-frozen.

But the rubicon has been crossed. "Reproductive Cloning: The Time is Near" is the headline of an editorial in the "Reproductive Biomedicine Online" journal published on the web on April 5th, 2003 – written by Zavos. "Recently out team of scientific and medical experts has created the first cloned human embryo for reproductive purposes", he writes. Zavos doesn't elaborate on the details, nor does he mention Illmensee's name. But what is clear to everybody: with this experiment the researchers have kicked open a door, and whether it possibly can be closed again remains to be seen. There is hardly any subject as controversial as the cloning of humans. It stands for the scary possibilities of gene technology, for hubris and delusional omnipotence of the researcher, for nightmarish science fiction scenery of genetically designed "Uebermenschen".

Critics of cloning advise against miscarriages and monstrous deformations, even against the horror vision of a genetically synchronized society. Only very few hail the opportunities of helping infertile couples to procreate with the help of cloned children and of multiplying humans with extraordinary abilities. The birth of clone sheep Dolly in early 1997 triggered worldwide discussions, her creators were denounced as breeders of modern Frankensteins. The "Spiegel" magazine even went so far as to print cloned Hitlers on its cover page. A lively debate on the ethical boundaries of biological sciences unfolded everywhere. Since Dolly researchers have cloned mice, rats, bovines, cats, dogs, pigs, horses, goats and even monkeys. But the cloning of humans for reproductive purposes remains to be a taboo.

Even the so called therapeutic cloning for the production of stem cells meant for medical purposes is heavily contended ethically, since cloned embryos are killed when extracting the stem cells. Reproductive cloning, that is, transferring cloned human embryos into the uterus, is even more disputed: it is outlawed worldwide, and in some countries even prohibited by law. But the state of legislation is inconsistent: The United Nations so far have failed to work out a joint resolution concerning human cloning. In many countries, like in Germany, reproductive and therapeutic cloning are prohibited. But there are other countries like Great Britain that place only reproductive cloning under punishment. The U.S. legislation is even more chaotic, with some states not even putting down any sort of regulation. In Kentucky, for instance, clone trials aren't liable to prosecution at all. In Germany, however, there are clear regulations defined in the legislation for the protection of embryos: "Anyone applying artificial methods to create a human embryo with identical genetic information as another embryo, fetus, human, or deceased person, is punishable with up to five years imprisonment or imposition of a fine."

But Illmensee is keen to retry cloning, he can't get over the fact that during his 2003 experiment the clone couldn't be transferred into the woman's uterus. In the beginning of 2004 he tries again, this time using the dermal cells of an infertile male and the egg cells of his wife. This time the embryo is successfully transferred, but no pregnancy develops. In 2005, there are more trials with three infertile husbands. I am surprised to see that Illmensee seems to consider breaking the taboo as some sort of leisurely competition: "Should I tinker with my motorcycles and vintage cars as a pensioner? I rather do a little work." Summing up he says: "Altogether we tried to clone five humans. One pair came from Egypt, two from the States, one from England, and one from another arabic country, Syria or Jordan. I was present when the egg cells were removed, and when the cloned embryos were transferred back into the uterus. Those were no donor or surplus oocytes – they were taken from the wives of the infertile husbands." According to him, nine cloned embryos were produced, one developed to a twelve-cell status before he was transferred into the uterus.

Why would a once internationally renowned researcher want to try to clone humans time and

time again? Why does he knowingly take the risk of miscarriages and deformations? And why does he engage into a task, which forces him to hide from the public? When Illmensee sets of as a researcher in the 1960s, the breath-taking progress of today's biotechnology seems yet unconceivable. The student of chemistry and biology sits in a lab at Ludwig Maximilian University in Munich, and concerns himself with the developmental genetics of fruit flies, Drosophila melanogaster. As he is still looking for a subject for his PhD thesis, he learns about the first successful cloning experiments on frogs. Why shouldn't one try the same with insects? Illmensee is hooked and doesn't let himself be deterred by the objection that there is no tool appropriate for the tiny fly eggs. For one year the PhD student fiddles about in order to design tiny glass pipettes. He works day and night – as if he was obsessed, say his friends of those days. "I can be quite persistent", Illmensee says. "Even if something didn't work out from the start, I wouldn't give up. I would keep on trying as long as it eventually works." Even Edison tried a thousand times until the electric light bulb shone a light, says he. Cooperating with the precision mechanics of Munich University, he assembles the necessary instruments piece by piece. And after a long series of trials he actually manages to remove the nucleus of Drosophila's egg cell and replace it by the nucleus of another embryo in a more advanced stage of development. But the manipulated eggs only divide a few times. Just one percent completes embryonic development so that a larva is hatched out. And only once a larva almost reaches the pupae stage. 40 years and innumerable cloning experiments later this result doesn't surprise him the least: In most cases less then one percent of cloning experiments are successful. Although Illmensee in 1968 fails to produce a cloned fly, his reputation as a researcher with "blessed hands" is established by an article in "Nature" magazine. Only in 2004 does a group of researchers from Canada manage to clone five flies – after 820 trials. During the 1970s, many labs are eager to hire the young man with the skill in fine technique. He moves to the U.S. and learns to manipulate the oocytes of mice. Then he receives the first offers for professorships, amongst others from Geneva University. There, the red carpet is rolled out for him: He receives more research funds and assistants than other professors. He has just about arrived in Geneva, when he is awarded the Marcel Benoist research prize – with a reward of 100,000 Francs the most lucrative scientific award Switzerland has to offer.

Equipped with everything a researcher yearns for, Illmensee in 1980 together with his US- colleague from the Jackson lab in Maine, Peter Hoppe, ventures into an attempt to clone mice. Parallel to his duties as a professor he mainly experiments during the nights and the weekends. His approach is different from that of the creators of clone sheep Dolly 16 years later: Illmensee takes fertilized oocytes, removes paternal and maternal genetic material and introduces the cell nucleus of a mouse embryo. After week and weeks of trials he manages to produce cloned mice embryos. Illmensee needs another 300 experiments to produce three cloned mice. In 1981, his paper is published in the renowned scientific journal "Cell" – and receives an enormous feedback from both scientific community and the public. Nobody had ever believed that cloning mammals might be possible at all – once again Illmensee's "blessed hands" had lead to a breakthrough.

At least that's what it first seems like. But behind Illmensee's back disaster is closing in: When he introduces a set of new experiments, two assistants of his cast doubt on the seriousness of the trials. A fact-finding-committee of Geneva University uncovers faults in the documentation of the experiments. Although allegations of forgery can't be proven the committee demands repetition of the experiments "in all scientific scrutiny", otherwise they were to be considered worthless. Although he has to deal with cuts of research funding, Illmensee manages to repeat the experiments. But then a study published in 1984 casts doubt over Illmensee's mice being cloned at all. For years no researcher had been able to repeat the cloning experiment. Many experts still believe in Illmensee possessing "blessed hands", but a scientific experiment is only valid if other scientists were able to repeat it. Therefore, the cloned mice are considered forgeries, and Illmensee an impostor. In 2006, researchers in Warsaw and one year later in Boston will eventually manage to show that mice can in fact be cloned using the method described by

Illmensee: by manipulating fertilized oocytes (zygote). This means Karl Illmensee could have been the first scientist to clone a mammal – 16 years ahead of Dolly's creators Ian Wilmut und Keith Campbell. But back then in the 1980s no-one believes him. The majority of professors in Geneva consider him a burden for the University's image. Illmensee doesn't want to be humiliated any longer he quits before the vote on the extension of his professorship, and accepts a post at Salzburg university. In Austria, he makes a living but lacks the financial means for top-notch research. While his family moves to Starnberg in the vicinity of Munich, Illmensee lives in a small flat in Salzburg – the decline seems foreseeable. He still can't cope with the accusation of forgery. He repeatedly asks his former employer in Geneva to rectify in public that the suspicion of forgery was never established. But nothing happens. The name Illmensee is slowly forgotten, scientific rehabilitation seems more and more unlikely. But when clone sheep Dolly puts the cat among the pigeons in 1997, Illmensee picks up courage again. He starts writing letters to old research buddies, corresponds with specialized magazines. But he still lacks the means to indulge into the research of cloning himself. Shortly after accepting a new position at Innsbruck gynecological hospital, Severino Antinori gets in touch with him in the beginning of 2001. The Italian reproduction expert became famous in 1992, when he helped a 62-year-old woman to have a child via donated egg cells in artificial insemination. Antinori wants to organize a conference on cloning in Rome, and he wants Illmensee to give a talk. The Austrian travels to Rome, and manages to establish contact to the cloning community – and only a few weeks later he is to meet Panayiotis Zavos.

The reproduction physicist Zavos is an enigmatic figure in science circles. He runs a hospital for artificial insemination in Lexington. Many think of the doctor as a loudmouth and a demagogue. On his website, Zavos celebrates himself as being a courageous pioneer. His motto: Being first is what really counts. The vision of his company Reprogen: Infertile couples shall be enabled to procreate via cloning. For years Zavos has made a fuss about announcing experiments. When it comes to cloning trials, his name keeps reappearing in the media. Illmensee remembers how he in cooperation with Zavos, Antinori and other physicists outlined a plan for cloning experiments on humans. But the project never came to be realized due to quarrels among the scientists. Afterwards, Zavos asked Illmensee whether the two of them shouldn't engage into "something serious" in Lexington. Illmensee travels to the small town in the States where he finds a "relatively well equipped lab". He is impressed by the "undeniable charisma" of his host. The urge to once again engage into clone research convinces him to stay. He becomes "Scientific Director" of Zavos' cloning company Reprogen. Cunningly, Zavos prepares a financial and institutional nest for the scientific outlaw Illmensee. Illmensee today admits that he was not lured into any kind of trap: "I could've said no after all", he says in our interview at Patras' beach. "I somehow slid into the whole thing. With his promise to make me director of a hospital in Limassol on Cyprus he made my mouth water." I ask Illmensee how he justifies his cloning experiments: "If you look at what all these infertile couples have been through – for me this is a good reason why one should try cloning", says he.

"Did you ever use the cells of dead people for cloning trials, as Zavos once stated at a press conference? For instance, the cells of an eleven-year-old girl who died in an accident?"
"We once unfroze the samples of the dead bodies and tried to cultivate them, but we never used them for cloning."
"You knew about the many mistrials in cloning other mammals, about the high risk of deformations of cloned embryos, and about the high probability for miscarriages. Why did you still pursue the cloning of humans?"
"In the beginning I thought cloning might be easier with humans than with other mammals. What do you know if something has never really been investigated? Only our work has shown how lousy cloning works with humans."
"But wasn't that foreseeable? What drove you to continue nevertheless?"
"It was, I have to admit, extreme curiosity. But when I saw the first results, I was convinced that

we shouldn't go on."
"Suppose it had worked out, would a clinical application be feasible at all?"
"No, because one needs approximately 200 donor egg cells and several transfers until a successful pregnancy and an infertile man getting a baby created by cloning. That's an incredible effort for only one patient. I don't see the slightest chance for clinical application, in my opinion, the project is failed."

On May 8th, 2007, Illmensee ends the cooperation with Zavos via Email. But the subject that has been occupying his mind all life long continues to move him – the curiosity can't be turned off. Today he cooperates with Mike Levanduski – not a dark horse in the field of cloning. The New- York-based embryologist cloned bovines with Mark Westhusin of Texas A&M University. In Levanduski's lab Illmensee deals with embryo splitting: After the first cell division he separates the two cells, so that both of them can develop into a complete embryo. Illmensee hence creates what is called monozygotic twins in nature – biologically speaking the same as clones. Embryo splitting may double the chances of successful pregnancy, says Illmensee, and, apart from that, one of the twins could be deep frozen to serve as a sort of stock for genetic spare parts for his living sibling. Surprisingly enough, the ethics commission of the American Society for Reproductiv Medicine considers embryo splitting a useful technique and finds research on it ethically acceptable. In Germany on the other hand, embryo splitting is prohibited and just like cloning ethically disputed, since the technique allows the creation of two or more genetically identical individuals.

40 years after his clone trials on flies, after his doubted cloning of mice, and the strongly criticized attempt to clone humans, Illmensee can't take his hands off the topic. Too strong is his inquiring mind, too big the curiosity, too much of a kick the feeling of being a discoverer. There is no doubt: Illmensee has long ago crossed an ethical boundary by transferring not one but nine clone embryos into the uteri of female patients.

The clone researcher Rudolf Jaenisch, who works at the Massachusetts Institute of Technology considers Illmensee's dealings with Zavos "a huge mistake". By doing so, Illmensee had done more severe damage to his scientific reputation than the accusations of forgery in the 1980s. And to lose his reputation is "the worst that can happen to a scientist". Apart from the personal drama of Karl Oskar Illmensee the whole story makes clear: The cloning of humans has already become reality. Concerning the technical equipment, it requires just no less than a middle-range IVF-lab such as Zavos'. And out of different motivations there will always be a demand – and sponsors, too. Equipped with a sufficient amount of oocytes, any other ambitious clone researcher could walk in Illmensee's footsteps and finish the task. This would very likely happen secretly, defying all social and institutional control. And one day, the first clone baby would smile from the front pages. Because no law has ever stopped the "extreme curiosity" of scientists like Illmensee: What can be done, will be done, one day. Vicarious agents like Zavos have the money and the unscrupulousness to attract the knowledge and talent of scientists like Illmensee.

I visit Zavos in Kentucky, a region of bavarian tranquility. Lush green and pretty farmhouses everywhere – and the neat little town Lexington lies in the midst of this idyll. A heavy door in glaring red color leads to Zavos' kingdom. Plush carpet floors, bombastic furniture in the living room. In his office the walls are covered by certificates, distinctions, and newspaper clippings. As a welcome the physicist, who once studied turkeys at Kentucky University, says in his Greek accent: "I am a pioneer, not a regular guy." He seems to be mighty proud of being involved in the cloning of humans. Proud enough to talk about where the experiments were conducted? "In our own lab", says he.

"Here in the U.S.?", I ask. Because that's what Illmensee stated, and one of his lab handwritten reports bears the headline "Lexington".
"No, no."

"In Cypres?"
"I can't tell you where. What I can't do here, I'll do elsewhere." He continues by listing several cities around the globe: "Damascus, Beirut, Kuwait-City, Shanghai, Peking ..."
"But how would you transport the equipment and the cells?"
"The kitchen goes where the cook is. We ship the stuff, and so on. We did clone human embryos and transfered them to uteri."
"Aren't you afraid of miscarriages, should a clone embryo ever be successfully bedded in the uterus."
"I don't think they would be abnormal. There are no studies on that topic. After implantation we would be observing the child by every conceivable method. Should it be deformed we could do what we do in IVF: abort."
"Many scientists think this attitude is unethical!"
"I probably have more enemies than friends, but I don't care."
"How do you get the funds for your cloning experiments?"
"A lot we pay for from our own pocket: maybe half a million, maybe two millions up to now. Then there are people contributing to our foundation - many people want these techniques to be advanced. And we charge minimal fees on those profiting from the techniques."
"But there are rules to science – some experiments ought not to be conducted despite the freedom of research. Especially not if human lives are put at risk."
"The egg cells we use stem from women who were informed on the experiments and who agreed to the proceedings, just as it is done in cases of IVF. Yes, we do take risks. Of course. But isn't crossing the street also taking a risk?"
"How will you continue your work, Mr. Zavos?"
"For instance, we are working on embryo splitting. By that we will create the first human with a spare wheel. Would you buy a car without a spare wheel? No, you wouldn't. One embryo grows to be a baby, the other is deepfrozen, and will become a source for embryonic stemcells later on. That is the future."
But who is to decide upon which embryo becomes a baby, and which the spare wheel?

Separate box: The trace leads to Germany
When running the first tests for his cloning experiments, Karl Oskar Illmensee replaced the precious human oocytes with bovine egg cells – whereinto he entered the cell nuclei of four infertile men. A functioning embryo cannot possibly evolve from such an inter-species clone, but the first stages of embryonic development proceed regularly. This makes the technique sufficient for studying the method, optimizing the conditions of the cell culture and for studying whether dermal, adipose or muscle cells are more suitable for the experiment. The method was officially approved in England to find out about the most favorable conditions for the production of cloned embryos for therapeutic purposes. In Germany, the production of inter-species clones is forbidden. "We created inter-species clones with the DNA of each of the infertile men and one woman that were willing to be cloned", says Illmensee. "We made a point that a cloning trial only made sense if these embryos developed favorably." Illmensee studied the technique of working with bovine oocytes and more cloning tricks with clone researcher Eckhard Wolf at his lab in Oberschleissheim, north of Munich. Another old colleague, veterinary geneticist Bertram Brenig of Goettingen University, analyzed the inter-species embryos – the costs being covered by Panayiotis Zavos. "We were not present, when the experiments themselves were conducted", says Brenig. "All we got were small plastic tubes with the cells. We were asked to check on what kind of DNA the cells contained." Illmensee stresses today that neither Brenig nor Wolf knew about the cloning experiments. "For me it is absolutely clear that reproductive cloning of humans is not acceptable", says Brenig. "I don't want to have anything to do with it." But then again: When the cells were analyzed in Goettingen, cloned embryos containing human DNA entered German territory. "But those cells were not viable", Brenig stresses. "Those were no clones any more, just a bunch of disintegrated cells." 

P.M.
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