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April 25, 2018

Welcome to the New Dawn of Cloning

Gone Are The Days of Boy Meets Girl

Clone Dogs

Reproduction used to be simple. Boy meets girl. Egg meets sperm. Nine months, one baby shower, and a whole lot of trips to Buy Buy Baby later, a new addition arrives to much billing and cooing by the proud grandparents. Oh those halcyon days. Today, the landscape is so much more nuanced as advances in technology enable procreation for individuals who would otherwise find it hard to conceive. Here at Cleanroom News, we’ve already drawn back the curtain on the world of in vitro fertilization (IVF) when we looked at cleanroom conception in ‘When Are Errors To Be Seen as Opportunities for Improvement?’ and have an on-going interest in the contamination control issues around human embryo fertilization in a cleanroom environment. But there is now another technology that has us concerned at the risk for contamination and it’s one that is only going to become more pervasive as the processes become more established and if – although that’s a big ‘If’ – the idea gains social acceptance. What are we talking about? Welcome to the world of cloning…

In 1932, English author Aldous Huxley published what has come to be considered a classic novel of dystopian futurism, Brave New World. In a tome frequently compared to and contrasted with George Orwell’s similarly futuristic dystopia in 1984, a eugenics-based society is maintained and controlled through the selective breeding of individuals created in laboratory conditions and indoctrinated to assimilate into pre-determined classes according to intelligence and labor needs. The ‘World State’ envisioned by Huxley is constructed according to the labor principles of Henry Ford’s assembly line, with homogeneity, mass production and consumption of replaceable goods, and predictability at its core. From the moment of conception, embryos are conditioned through chemical, thermal, and environmental stimuli and, upon birth, are programmed to believe their class to be superior and the others merely necessary for society to function. Individual discontent or malaise is eradicated through the use of anti-depressant medications and a mind-altering drug known prosaically as ‘soma.’ The only elements missing in Huxley’s vision of a future controlled and manipulated within the laboratory are genetic engineering (the structure of DNA had not then been discovered) and the concept of cloning.

‘How afraid of human cloning should we be?’

Despite the relative success of the creation of Dolly the Sheep by Scottish scientists back in 1996, the science of producing exact replicas of an individual is still somewhat patchy. Several mammals have since been cloned – cats and pigs, for example – but human cloning is (some say mercifully) still some years away. But that said, in January of this year Phillip Ball of London’s The Guardian penned an interesting op-ed on the extent to which we, as a society, should be mindful of the reach of the clones.(1) In ‘How afraid of human cloning should we be?’ Ball examines how the recent cloning of macaque monkeys in China seems to bring human cloning one step closer. Successfully creating primates has always been seen as the holy grail of this branch of genetic research, as primates – like ourselves – are notoriously difficult to reproduce. Despite outlandish and unsubstantiated claims by the Raelians, a fringe organization headquartered in Geneva, Switzerland, to have cloned a human baby in 2002, no human clone has ever been proved to have been successfully generated to date. But the cloning of the macaque monkeys was significant insofar as it was achieved by refining the process used in the creation of Dolly. By taking the cells of a fetus, as opposed to an adult donor, scientists were able to ‘awaken’ genes crucial to development of the embryo that are not revived when using adult donor cells. Of the 79 cloned embryos transplanted into 21 surrogate mothers, only 6 resulted in pregnancy, of which the baby macaques Hua Hua and Zhong Zhong were the only live births. Perhaps a disappointing success level.

…‘advanced biotechnology for industrial and biomedical applications using animal cloning and pluripotent stem cells combined with transgenic technology.’

But in the world of canine cloning, if results from South Korea are anything to go by, the future looks more promising. Until recently, dog lovers among us were forced to face the stark truth that our four-legged companions would be with us only for a short time. With life expectancies of 7 to 17 years depending on breed, our dogs are fairly certain to predecease us, a fact of life that can be incredibly hard to overcome. So it is understandable that some people will go to arguably extreme lengths to recapture that which they have lost. And, given sufficient funding and commitment, that can mean more that simply finding another dog of the same breed, with similar features. With today’s technology, it can mean the potential of cloning your companion and having those years all over again. Sooam Biotech Research Foundation (SBRF), based in Seoul, South Korea, is a non-profit organization that focuses on ‘advanced biotechnology for industrial and biomedical applications using animal cloning and pluripotent stem cells combined with transgenic technology.’ Quite the mouthful. Founded in 2006, scientists at the organization cloned their first canine – Missy – a year later, achieving international recognition in 2012 ands 2013 with their inclusion in documentaries by both National Geographic and TLC/Discovery channel. According to the company’s website, it also worked with the National Police Agency in 2012 to clone ‘special purpose dogs’ although further details of this endeavor are not presented.

The tools used to punch each sample from the skin or muscle must be cleaned with alcohol between tissue collections…

To date, SBRF claims to have cloned 600 companion animals using technology ‘for any dog no matter its age, size, and breed […healing] broken hearts.’(2) And despite the hefty price tag, the process seems remarkably straightforward. For $100,000, SBRF can clone your pet whether alive or dead. Instructions on the website guide both the pet owner and their veterinarian through the process of harvesting an aseptic sample that can be shipped to South Korea. In the case of a deceased individual, 6 to 8 vials of biopsy material must be obtained within five days of death, from an area of the body that has first been shaved and cleaned with a series of betadine, chlorhexadine, and alcohol scrubs. For a deceased animal, the biopsy samples should be composed of skin tissue (epidermis, dermis, and parts of the subcutis) and muscle tissue, with each ‘punch’ being 8mm in diameter, or around the size of a nickel. The tools used to punch each sample from the skin or muscle must be cleaned with alcohol between tissue collections, and the materials preserved in vials containing sterile saline solution. Each vial should be sealed with Parafilm Stealing Strip and maintained at 35° to 41°F until transported to the company.

So far, so grizzly but, for a bereaved pet owner with $100k to spend, perhaps it is a logical step. Shipped via carrier, the sample clears Customs in Korea and, once in the lab and determined to be viable, is sterilized and separated into multiple pieces. Treated with a reagent, the cells undergo a chemical dissociation as they separate from the tissue. Using a centrifuge, the cells are harvested for culturing in a growth medium and 7 to 14 days later, the cloning – and the controversy – can begin.

SBRF rents two dogs from a Laboratory Animal Provider service – one to be an egg donor and the other a surrogate mother to the clone. After a course of fertility treatments to encourage egg development, the donor dog is ‘flushed’ – that is, her eggs are collected surgically and their DNA is removed in a process known as enucleation. The emptied egg then has no genetic material associated with the donor and is ready to receive that of the dog to be cloned.

…in the case of cloning, the sperm is replaced by a combination of additional skin cells treated with an Electro Cell Manipulator…

So now we have an egg but, thinking back to our Biology 101 class, we also need sperm to create a baby, correct? No, in the case of cloning, the sperm is replaced by a combination of additional skin cells treated with an Electro Cell Manipulator, a generator that stimulates the egg and skin cells electrically, fusing them together to create a fertilized embryo. After just one minute, a batch of up to 15 embryos is created, which can be transplanted into the surrogate mother. After 30 days, testing is performed to verify pregnancy and in a further 30 days delivery of a live puppy may occur. We say ‘may’ as the success rate claimed by the company stands at 40%, although if no viable pregnancy results on the first try, a second surrogate mother will be used. In the event of a successful live birth, SBRF will care for the puppy until the owner arrives to claim it or will transport to the owner using a chaperone.

Barbra Streisand, for instance, famously commissioned clones of her Coton de Tulear dog, Samantha.

And who are these potential owners? If the headlines are anything to go by some Hollywood glitterati are very interested in ways in which to keep their pets alive, even beyond death. Barbra Streisand, for instance, famously commissioned clones of her Coton de Tulear dog, Samantha. While they resemble their genetic progenitor in appearance, the resulting pups, Miss Scarlett and Miss Violet, do not have the same personality, a cause of some chagrin to Ms Streisand. And then there’s the case of the 911 ‘hero dog,’ Trakr, a German Shepherd who was instrumental in rescuing the last survivor of the World Trade Center attack in 2001. Although he succumbed in 2009 to a neuro-degenerative disorder as the result of his work in the rubble of the towers, Trakr was survived by five clones – Solace, Valor, Prodigy, Trustt, and Dejavu – created by SBRF.

Unlike the human egg donors and surrogate mothers, canines used in the creation of clones neither consent nor are paid for their efforts.

However, cloning is not all about feel-good stories of healing hearts by replacing lost pets and heroes. Although the technology may be exciting for the bereaved pet owner, the process is inevitably controversial. In an interesting video report by TechInsider, ‘The Science Behind Dog Cloning,’ the narrator does acknowledge that while the desire to create a clone of a much loved pet is understandable, and the solution is available for a price, the decision to create a clone is not ‘ethically uncomplicated.’(3) Unlike the human egg donors and surrogate mothers, canines used in the creation of clones neither consent nor are paid for their efforts. Unlike the ‘gestational carrier’ – Kim Kardashian’s preferred term for ‘surrogate’ – employed by the reality TV star and her husband Kanye West, reportedly to the tune of $180,000, the rented bodies of the dogs are sourced from a pool of candidates who have been purpose bred solely for research.(4) And, as with any ‘farmed’ animal, their needs and wishes are rarely accorded more than minimal consideration by those with a vested fiscal interest in their use.

As with IVF processes, cloning must be done in a sterile environment.

And, of course, in addition to the thrill of the pure science, profitability is a driving factor for industry players. And where the pursuit of profit regulates innovation, shortcuts may be the result, allowing for the risk of contamination. As with IVF processes, cloning must be done in a sterile environment. In any situation in which polymerase chain reaction (PCR) is used to make copies of a region of DNA, the cleanroom should have two separated areas – one for pre-PCR work, and the other for post-PRC tasks. The two independent airflows (one for each area) should be unidirectional and regulated by HEPA filtration, and it almost goes without saying that consumables used in one area should never be introduced into the other. Gloves, papers, PPEs, masks etc. should be sourced from each area’s dedicated supply, and gloves especially should be changed frequently. Aseptic cleaning techniques should be applied, per the laboratory’s standard operating procedures (SOPs), to all surfaces and touch points, and wipe tests should be conducted periodically to proactively monitor contamination levels and head off problems before they occur. After all, in the genetic research field, the ‘end product’ – which can be as easily damaged as any microchip on an assembly line – is destined to be a living, breathing, sentient being.

In many ways, cloning is both a nascent and an ancient concept. As the U. S. Food and Drug Administration (FDA) notes on its website ‘we eat fruit from plant clones all the time, in the form of bananas and grafted fruits. We’ve been cloning plants for decades, except that we refer to it as “vegetative propagation.”’(5) And in nature, some animals – from the hammerhead shark to the komodo dragon, the starfish to the gall wasp – can, under the right conditions, clone themselves. Sometimes, as in the case of the ‘virgin birth’ of a hammerhead shark pup in Henry Doorly Zoo, Omaha in 2007, it is a case of parthogenesis and at other times, as in the case of the gall wasps, it is simple asexual reproduction of unfertilized eggs that mature into clones. To date, the only concern of our regulatory bodies has been the caution that the flesh and secretions (meat and milk) of cloned farmed animals must not enter the human food chain. However, the agency has relied upon clone producers and breeders to abide voluntarily upon the ban and says that ‘To the best of our knowledge, they have done so.’(6) Well, that’s reassuring then.

In his 1985 treatise Amusing Ourselves to Death: Public Discourse in the Age of Show Business, Neil Postman highlights the opposing contentions in Orwell’s 1984 and Huxley’s Brave New World, two novels in which the future is controlled through manipulation, fear, science and advanced technologies, and social misdirection. Postman comments that ‘Orwell feared those who would deprive us of information. Huxley feared those who would give us so much that we would be reduced to passivity and egotism. Orwell feared that the truth would be concealed from us. Huxley feared the truth would be drowned in a sea of irrelevance. Orwell feared we would become a captive culture. Huxley feared we would become a trivial culture, preoccupied with some equivalent of the feelies, the orgy porgy, and the centrifugal bumblepuppy. As Huxley remarked in Brave New World Revisited, the civil libertarians and rationalists who are ever on the alert to oppose tyranny “failed to take into account man’s almost infinite appetite for distractions.”’(7)

In the present era of always-on, 24/7, internet-mediated information overload, celebrity gossip, Insta-Reddit-Facebooked-Tweeted-Bloggered-viral-YouTubed sensationalist pabulum, fragmented attention spans, and virtually limitless choice, Postman’s apocryphal warning that technology would enable an abundance that would result in social enslavement seems ever more relevant. And the visions of an engineered society of which both Orwell and Huxley warned ever closer. And cloning – the laboratory creation of individuals selected for distinct physical or intellectual characteristics – could well be a significant part of that future. Could it be that, while we squander our time frolicking in the news shallows of the latest reality TV star’s surrogate pregnancy or the aging Hollywood diva’s cloned lapdogs, we fail to notice the encroachment of technology upon our free future? As we become so distracted that we do not even notice we are drowning in push notifications on our social media feeds, the technological advances developed to recreate man’s best friend could, if unchecked, go on to be applied to the creation of man himself? A brave new world indeed…

Would you consider cloning your best friend? Do you have $100k to spare? If so, we’d love to hear from you!

References

  1. https://www.theguardian.com/commentisfree/2018/jan/25/afraid-cloning-humans-macaque-monkeys-reality
  2. http://en.sooam.com/dogcn/sub01.html
  3. https://www.youtube.com/watch?v=DmHYUvmiXQI
  4. http://fortune.com/2017/09/06/kim-kardashian-pregnant-surrogate/
  5. https://www.fda.gov/AnimalVeterinary/SafetyHealth/AnimalCloning/ucm055512.htm
  6. ibid
  7. Postman, Neil; Amusing Ourselves to Death: Public Discourse in the Age of Show Business (London, Penguin Books, 1985).
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