When George Orwell penned perhaps his most famous piece of fiction back in the 1940s, the English author and journalist foretold a revolutionary and futuristic 1984; an era of totalitarian regime, public mind control, and groundbreaking concepts.

As a social science fiction, Nineteen Eighty-Four would go on to become an iconic novel and cultural phenomenon.
But even the literary works of this social commentator and visionary couldn’t foresee the moment of marvel that would actually take place in 1984.

For March 1984 opened a whole new chapter in medical history, as the first baby born from a liquid nitrogen-frozen embryo came into the world. Baby Zoe was born in Melbourne, Australia and quite apart from the wonder of another new life on Earth, heralded a significant milestone in the field of cryobiology.

Now more than 25 years later and in a fast-paced modern society where around three million babies have been born through associated reproduction techniques, it seems fitting to reflect on a day when a few litres of liquid nitrogen ushered in a brave new world.

But in truth, the birth of Zoe is just one branch of the complex cryobiology sector. And the concept of a ‘test-tube baby’ wasn’t new either; Louise Brown had become the first ‘fresh test-tube baby’ in England six years before. What was so radical, was the experimentation with a frozen embryo, and the awareness that would follow.

What is cryobiology?
A branch of biology, the field of cryobiology is the study of cold life sciences or life at low temperatures – quite literally.

Derived from the Greek words cryo, bios and logos, these translate as cold, life, and science. That might be simple enough, but the actual business of cryobiology itself is both complex and diverse. It’s important to point out that cryobiology includes or is associated with several distinctions or major areas of study which include, but are certainly not restricted to, cryogenics, cryonics, and cryopreservation.

Cryonics for example, focuses on the low temperature preservation of humans and/or mammals – with the intention of future revival – and is not generally considered to be part of the mainstream cryobiology sector. Cryopreservation meanwhile, is the technology of preserving cells, embryos or whole tissues at ultra-low temperatures, often for medical purposes and in human science, though also applied in the animal world and nature itself.

It’s not uncommon in nature that plants and other organisms survive weeks and months in a preserved, frozen state, as many living organisms are able to tolerate extensive spells under temperatures below the freezing point of water. While these subjects are usually able to accumulate ‘natural’ cryoprotectants such as anti-nucleating proteins and polyols to guard against frost damage by ice crystals, in human science this is something provided artificially.

With such an array of applications across science and nature and in the worlds of both humans and animals, it’s easy to see how cryobiology has become so significant in modern life. It’s big business and while it may be difficult to place an exact value or financial worth on the overall cryobiology market, this represents a substantial and fast-growing segment of the healthcare sector.

So much so, that for the industrial gas community alone, cryobiology is emerging as an important market. This was highlighted in summer 2009, when Air Liquide’s Healthcare business acquired California-based Pacific Science, Inc. – a provider of cryobiology equipment and services to biobanks.
Pacific Science has grown rapidly since its inception in 1995, in line with the boom in biotechnology.

For Air Liquide, the acquisition was not only consistent with the group’s Healthcare business development in the US, but also affords an added route into one of the most rapidly growing segments in healthcare.

North America is thought to represent more than 40% of this key market. Coupled with other subsidiaries such as its Canada-based VitalAire firm, Pacific Science gives Air Liquide a channel through which its cryobiology solutions can be brought to North America – including its designs for biobanks, vacuum line installations, liquid nitrogen supply, and freezing & storage capacity.

Of course many other industrial gas and equipment companies like Taylor Wharton International, for example, already cater for the cryobiology end-users. But when a Tier 1 major player like Air Liquide shores up its interest in the market, this is a further reflection of the confidence held in this area.

Fellow Tier 1 player The Linde Group is just as effusive in its interest for the cryobio industry. Shivan Ahamparam, Market Segment Manager at Linde told gasworld, “We believe that biotechnology is a vital component of the life sciences market and requires manufacturing processes that are different from those of traditional pharmaceutical drugs.”

“It is critical to control the microenvironment of these highly complex protein-based formulations at every stage of the product life cycle so that they maintain desired quality. This often requires cryogenic freezing methods.”

“Linde believes,” Ahamparam adds, “the industrial gas industry, with its greater expertise in this field, can help design applications to meet this need.”

Significance in modern society
Vast, diverse and still relatively in its youth, we’ve already established the growing importance of cryobiology in the fabric of modern society.

This is especially relevant and perhaps no more newsworthy than in the area of medical research and medical advancement. Cryopreservation is central to this; from the many organs, tissues and cells routinely stored and transported at low temperatures for short periods, to cell suspensions like blood and semen and thin tissue samples often stored almost indefinitely in liquid nitrogen.

Human gametes (sperm, eggs) and embryos are now routinely stored in fertility research labs, where controlled-rate and slow freezing processes are fundamental.

This was the case with that breakthrough birth of Zoe in 1984, when Zoe’s mother had produced 11 eggs, or oocytes, which were fertilised and then frozen using a controlled rate freezer manufactured by London (UK) company Planer plc. Frozen in liquid nitrogen, the embryo was later thawed and implanted – a technique which has become commonplace by the 21st century.

In fact, the number of live births from so-called ‘slow frozen’ embryos is believed to be circa 500,000 or around 20% of the estimated three million IVF (in vitro fertilisation) births worldwide.

And so, cryopreservation – and cryobiology – is becoming so fundamental to modern society.

The preservation of biological matter is increasingly important, because human and animal tissue needs to be stored for research and regenerative medicine purposes: bone marrow for cancer treatment; blood products for collection of stem cells and transfusion; sperm, eggs, embryos for fertility; skin, bone and cell lines for transplantation; plant cells, algae cultures, protozoa, seeds and fungi for the purposes of sustainable agriculture, biotechnology and conservation and biodiversity.

The deep cold of liquid nitrogen is needed to stop any biological action and using mechanical type refrigerators, even very cold ones, is not feasible. Many of the cells needed for medicine are difficult to freeze and are frozen down at a predefined protocol, preventing damage to cells before they are then transferred to liquid nitrogen storage tanks.

Cryogenic storage at very low temperatures (from the -80°C down to -196°C range) is presumed to provide an indefinite, if not near infinite, longevity to cells – although the actual ‘shelf life’ is rather difficult to prove. However, it is suggested that human gametes and certain embryos can actually survive cryopreservation at -196°C for decades under well-controlled laboratory conditions.

Scientific researchers’ attention is also now turning towards the preservation of other, often larger, samples, which are currently impossible to successfully freeze and thaw – such as cartilage. If research and testing is successful, perhaps one day even whole organs might be preserved using freezing gases, for later transplantation.

A new era?
A future application that’s already being realised is the use of umbilical cord stem cells.

Stem cell research is not without controversy or detractors, with the ethical debate largely focused on embryonic stem cells. Those in favour will point to the fact that stem cells can potentially repair extensive tissue damage and further still, embryonic stem cells can become all cell types of the body due to their pluripotent nature.

However, those against highlight how current technology limitations mean that a human embryo must first be destructed to create a human embryonic stem cell; this is seen as violating the ‘sanctity of life’.

Yet a whole new era of stem cell research and usage could be just around the corner if cord blood is anything to go by. Stem cells are characterised by the ability to renew themselves through mitotic cell division and differentiate into a diverse range of specialised cell types – hence described as pluripotent. If these incredible qualities can be harnessed via a non-controversial source, it could usher in another breakthrough in human medical history.

Enter, cord blood. It might be commercial, and there is an element of stigma attached from certain detractors, but those in favour of cord blood see this as a revolutionary new field.

Until recently, umbilical cord blood was generally discarded (after childbirth) with the umbilical cord itself and the placenta. By simply saving this and cryopreserving it at -196°C in a liquid nitrogen tank, however, a stem cell-rich source becomes available.

The success of cord blood transplantation does of course depend on tissue compatibility, but according to US-based HealthBanks Biotech Company PacifiCord, cord blood stem cells are used to treat over 70 diseases and can be used to treat leukemia, sickle cell disease, and metabolic disorders.

It’s thought that this could potentially extend to heart disease, muscular dystrophy, diabetes, Alzheimers and more.

PacifiCord describes stem cells as the ‘building blocks underlying a new era in medicine’. We wonder, could Orwell have imagined a future era so built on the foundations of cryobiology?