Researchers Grow Stem Cells Without Direct Use of Animal Cells

In March, researchers at the Roslin Institute in Scotland reported they had managed to grow stem cells without directly exposing the stem cells to animal cells, though the stem cells were still indirectly exposed to animal cells.

Until now, embryonic stem cells are grown using mouse feeder cells and in animal growth serum. There is some concern that as long as animal cells are used in the process that there is a risk that an animal virus will be transmitted to human beings who might someday receive stem cell transplants.

The Roslin Institute researchers used human protein laminin and feeder cells taken from human neonatal foreskin cells to provide the nutrients that the embryonic stem cells need.

This process is still not entirely animal-free, however, as the neonatal human foreskin cells are themselves grown in a process which uses animal cells.

Almost certainly at some point embryonic stem cells will be grown end to end in non-animal substrates at which point the animal rights movement will claim that animal research had nothing at all to do with embryonic stem cell research.


Human embryonic stem cells grown animal-free. Andy Coghlan, New Scientist, March 17, 2005.

‘Animal free’ stem cells created. The BBC, March 16, 2005.

PCRM Develops Animal-Free Insulin Assay

Physicians Committee for Responsible Medicine reported in February that it had worked with BiosPacific and Linco Research to develop an insulin assay that measures insulin levels in individuals without relying on animal products.

PCRM president Neal Barnard said in a press release that PCRM needed to conduct insulin assays as part of a study of the effects of vegan diets on type 2 diabetes. Barnard said,

We only had two options available to us when we began our diabetes trials. One, we could use test kits with insulin antibodies grown in vivo — literally from cells injected into the abdomens of live mice — or we could use kits containing antibodies from cells cultured with fetal calf serum. Neither was acceptable to us.

So PCRM’s Megha Even worked with California lab BiosPacific to create an animal-free replacement for the fetal calf serum, and then with Linco Research created a test that uses antibodies cultured in the non-animal serum.

According to Barnard, details of PCRM’s non-animal assay will soon be published in a peer-reviewed journal, and the non-animal test will be made available commercially,

We hope that by making the test readily available and competitively priced, researchers and medical labs will use it. We have proven that if researchers are willing to make the effort, there are effective, humane alternatives to animal-based assay, and other testing procedures — alternatives that could help save the lives of millions of people and animals.


PCRM develops world’s first cruelty-free insulin assay. Press Release, Physicians Committee for Responsible Medicine, February 9, 2005.

Indian Researchers Develop Software to Assist in Drug Development

The BBC reports that Indian researchers have created computer modeling software to allow researchers there pre-screen potential drug compounds before moving to animal tests with the compounds.

Such systems are routinely used in the United States, but such software tends to be expensive and out of reach of the developing world.

Developed by Delhi’s Invenio Biosolutions, the Drug Discovery Assistant is currently being used by India’s Central Drug Research Institute. According to the BBC, the program will help reduce the number of trials involving animals from 20-30 to 2-3 in the initial evaluation of a compound.

The upshot of this — and the reason such systems are used widely in the developed world — is this will cut in half the drug development time (which, according to the BBC, is currently around 20-25 years in India).


Indian software replaces lab animals. Ram Dutt Tripathi, The BBC, June 6, 2003.

Researchers Explore Ways to Reduce Animals Killed in Environmental Testing

Scientists at the University of Georgia’s Savannah River Ecology Laboratory recently reported on a new toxicology study that might reduce the number of animals killed to test how much of an environmental contaminant remains in animals living in environments where the contaminant is present.

Traditionally animals are captured, killed and then tested for contaminant levels. University of Georgia researchers Brian Jackson, William Hopkins, and Jennifer Baionno are working on an alternative that involves using a laser to take small samples from the tails of animals rather than killing the animal.

They took a group of banded water snakes, and fed two groups of snakes contaminated fish (they also used a control group that was fed non-contaminated fished). The researchers then used the laser technique on one of the groups of snakes and the traditional method on the other group of snakes, and then compared the resulting data.

The similarity was close enough that they concluded that,

Taken together, the findings from this study suggest that laser ablation of micro-dissected tissue shows promise as a non-destructive technique for conservation-minded exo-toxicological studies.


UGA scientists test less lethal means to determine contaminant uptake. Press Release, University of Georgia, July 21, 2003.

In Defense of Animals/Fund for Animals Claim Victory that Wasn't

At the In Defense of Animals web site, IDA reprints a list of “Animal Rights Victories in 1999” that was compiled by Michael Markarian of The Fund for Animals. Not surprisingly, one of the “victories” on the list never actually happened. Midway through the list is this item,

The NIH banned the use of mice in monoclonal antibody production, saving the lives of up to one million mice per year, and admitting that animals feel “pain, distress, or discomfort.”

The only problem is that the NIH did not ban the use of mice to produce monoclonal antibodies and the reason it decided against a ban goes to the heart of the debate over animal research.

What’s a monoclonal antibody? It is a method of mass producing specific antibodies. Researchers take tumor cells that will reproduce forever if given the proper nutrients and fuse those with cells that produce specific antibodies. The result is called a hybridoma which is then cloned to produce large numbers of cells that will produce specific antibodies.

The ability to produce monoclonal antibodies is a direct result of years of animal research and animals are essential for the first phase of the process, the creation of a hybridoma. Typically, a hybridoma is created by immunizing an animal (almost always a mouse), and then obtaining immune cells from the animal’s spleen. These cells then get fused with the tumorous cancer cell so that they can reproduce indefinitely.

Nobody suggests that there is an animal alternative to this process. Regardless of how they are later cultivated, monoclonal antibodies require the use of an animal during the initial phase.

But cultivation of these cells is another story. Researchers are able to growth monoclonal antibodies either in vivo or in vitro.

The in vivo model involves injecting animals (again, almost always mice) with hybridomas. The hybridomas reproduce and produce a fluid called ascites on the animal’s abdomen. The fluid contains a large number of monoclonal antibodies that can then be harvested for further study.

The in vitro model involves culturing the hybridomas in one or another culturing medium. Note that this also involves the use of animals (though not whole animals), with the most popular method of culturing being using fetal bovine serum.

In 1997, the American Anti-Vivisection Society petitioned the National Institutes of Health to prohibit researchers receiving NIH grants from using the whole mouse method to produce monoclonal antibodies. Since in vitro methods were available to produce the antibodies, AAVS argued, animals were suffering needlessly.

Contrary to what IDA apparently believes, the NIH rejected an outright ban. Instead, after commissioning a study of the issue from the National Research Council, it issued a policy that for NIH grants in vitro methods of monoclonal antibody production should be the preferred method of production.

Using whole mice to produce monoclonal antibodies is still allowable under NIH, however, it in vitro methods are not suitable for one reason or another.

The National Research Council that looked into the issue found that there is a continued scientific need to produce monoclonal antibodies in mice. According to its 1999 report (which is available here),

There are several reasons why the mouse method of producing mAb cannot be abandoned: some cell lines do not adapt well to tissue-culture conditions; in applications where several different mouse mAb at high concentrations are required for injection into mice, the in vitro method can be inefficient; rat cell lines usually do not efficiently generate mAb in rats and adapt poorly to tissue-culture conditions but do produce mAb in immunocompromised mice; downstream purification or concentration from in vitro systems can lead to protein denaturation and decreased antibody activity; tissue-culture methods can yield mAb that do not reflect the normal modification of proteins with sugars, and this abnormality might influence binding capacity and other critical biologic functions of mAb; contamination of valuable cell lines with fungi or bacteria requires prompt passage through a mouse to save the cell line; and inability of some cell lines that do adapt to tissue-culture conditions to maintain adequate production of mAb poses a serious problem. For these reasons, the committee concludes that there is a scientific necessity to permit the continuation of the mouse ascites method of producing mAb. However, note that over time, as in vitro methods improve, the need for the mouse ascites method will decrease.

Maybe someday there will be no need to use mice to mass produce monoclonal antibodies, but that day is not yet upon us.

At the time the NIH changed its policy, the American Anti-Vivisection Society estimated that about 90 percent of monoclonal antibody production done as part of NIH grants would move to in vitro models, with the other 10 percent still being performed in vivo. So far, this writer is unaware of any research on just how much the new policy affected the landscape of monoclonal antibody production.

But, one thing that did not happen was an outright ban of antibody production in mice as In Defense of Animals and The Fund for Animals claimed.


Animal Rights Victories in 1999, Compiled by Michael Markarian of The Fund for Animals For the 2000 Summit for Animals. In Defense of Animals, 2000.

Animal protection group precipitates historic policy change at NIH. Press Release, American Anti-Vivisection Society, December 22, 1999.

Monoclonal Antibody Production. National Research Council, 1999.

The Promises and Limitations of Animal Alternatives in Medical Research

In 1997 an alternative to animal testing for certain skin tests hit the market. The procedure — which uses lab-grown human skin samples — is beginning to increase in popularity with sales of the product growing by 40 percent per year.

On the one hand, this sort of in vitro test has a number of advantages to whole animal tests. They tend to be cheaper and faster than in vivo animal models. Animal rights activists like to pretend that animal research persists largely for economic reasons, but in fact any animal alternative that speeds the time it takes to get a product to market will find a ready market.

On the other hand, unlike the animal rights activists, those who develop and use such animal alternatives also recognize that they have inherent limitations. Technology Review quotes Charles Hewitt, director of surgical research for Robert Wood Johnson Medical School, noting that although he uses the bioengineered skin in his research, “We can’t get all the responses we need to test just from our model.”

In fact, as Technology Review notes, often these “animal” alternatives in fact end up being alternatives to traditional human clinical trials. A major use of bio-engineered skin, for example, is to screen compounds before sending them to human clinical trials.

Researchers at the State University of New York at Stony Brook, for example, evaluate drug under consideration for nasal use. They used to send such drugs directly to clinical trials, but now first use tests on the bio-engineered skin to screen out drugs likely to fail in clinical testing, so that only the most promising drug candidates are subjected to expensive clinical testing in human beings.

Animal rights activist claim that all animal tests could be replaced with alternatives tomorrow, but the reality is that a mix of human and animal, in vitro and in vivo tests and experiments will form the foundation of medical research for the forseeable future.


Saving skin. Alan Joch, Technology Review, February 11, 2002.