Sel Penyebab Leukemia Ditemukan

Selasa, 29 Januari 2008 | 13:28 WIB

TEMPO Interaktif, Oxford:
Tim peneliti menemukan bahwa kedua anak kembar tersebut memiliki sel tunas abnormal praleukemia dalam darah mereka. Sel itu bisa "tidur" dalam sumsum tulang atau berkembang menjadi sel tunas leukemia. Hasil ini dikonfirmasi oleh eksperimen yang menggunakan sel tali pusar manusia.

"Penelitian ini berarti kami dapat mengetes apakah penanganan leukemia lymphoblastic akut pada anak bisa dikaitkan dengan menghilangnya dan berkembangnya sel tunas leukemia," kata Profesor Tariq Enver dari Unit Hematologi Molekuler Universitas Oxford, yang memimpin penelitian tersebut. "Mulai saat ini, upaya penyembuhan bisa difokuskan pada upaya membidik sel tunas praleukemia dan sel tunas kanker dengan obat yang ada atau yang akan kita kembangkan."

Upaya penyembuhan yang terfokus, menurut Tariq, bisa menghindari efek samping pengobatan kanker kemoterapi yang menyakitkan dan terkadang justru membahayakan kondisi tubuh pasien. Hal ini sangat penting karena terbukti, Olivia, salah satu anak kembar yang terkena leukemia, mengalami kebutaan di sebelah matanya akibat infeksi yang tidak bisa dilawan tubuhnya saat kemoterapi.

Para ilmuwan telah melacak kemungkinan sel tunas prakanker itu akibat fusi abnormal dari dua gen yang terjadi selama kehamilan ibu. Fusi ini menghasilkan protein hibrida, sebuah "kesalahan" genetik yang terjadi secara acak dan menyebabkan sel menjadi terjangkit leukemia. Gen yang diambil dari si kembar lantas ditransplantasikan ke tikus laboratorium yang mengkonfirmasi adanya hubungan langsung antara malfungsi genetik dari sel tunas tersebut dan leukemia.

Lembaga donor Inggris yang membiayai penelitian itu, Leukemia Research and the Medical Research Council, dan Rumah Sakit Great Ormond Street menyatakan sangat gembira atas penemuan itu dan berharap penelitian dilanjutkan ke upaya mencegah dan mengobati penyakit tersebut.

AMAL IHSAN | SCIENCEDAILY

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Next Steps for Stem Cells

By Emily Singer

Searching the brain of an Alzheimer's patient for clues into the origin of the disease is like trying to find the cause of a plane crash in the wrecked aftermath. However, a recent breakthrough in stem-cell research could generate new cellular models that allow scientists to study disease with unprecedented accuracy, from its earliest inception to a cell's final biochemical demise.

Last November, two groups of scientists announced that they had independently achieved one of the stem-cell field's biggest goals: the ability to reprogram adult cells into embryonic-like stem cells without the need for human embryos. (See "Stem Cells without the Embryos.") The findings garnered extensive media attention, largely because the new method obviated the need for human embryos, a major ethical minefield that has stymied research.

But scientists at stem-cell labs around the world are excited for another reason. The technique creates cells that are genetically matched to an individual, meaning that it's now possible to create novel cell models that capture all the genetic quirks of complex diseases. "Being able to have human cells with human disease in a dish accessible for testing is a real boon to technology and to science," says Evan Snyder, director of the Stem Cells and Regeneration Program at the Burnham Institute, in La Jolla, CA.

While animal models exist for many human diseases, they typically only incorporate certain aspects of the disease and can't capture the complexity of human biology. In addition, some disorders known to have a significant genetic component, such as autism, have proved difficult to model in animals.

To reprogram cells, scientists from Wisconsin and Japan independently engineered skin cells to express four different genes known to be expressed in the developing embryo. For reasons not yet clear to scientists, this treatment turns back the developmental clock. The resulting cells are pluripotent, meaning that they can develop into any type of cell in the body, and they can apparently divide indefinitely in their undifferentiated state. The first two published studies on the new technique reprogrammed cells from a skin-cell line, while a third study, published last month, generated stem cells from the skin biopsy of a healthy volunteer.

No one has yet generated cell lines from a patient, although scientists have been talking about doing so for years. Previously, the only way to make such models for complex genetic diseases was through human therapeutic cloning, also known as nuclear transfer, which is fraught with technical and ethical issues and has not yet been achieved. (See "Stem Cells Reborn" and "The Real Stem Cell Hope.") "Assuming that these procedures are as easy to do as it seems, it's definitely more tractable than nuclear transfer," says Snyder. His own lab is trying to generate such models, as is "probably everyone else you could call on your rolodex," he says.

To generate a disease-specific cell model, scientists would take some cells from a patient with a particular disease and revert them to an embryonic state. The cells would then be prodded to develop into the tissue type damaged in that disease, such as dopamine neurons in Parkinson's disease or blood cells in sickle-cell anemia. By comparing the differentiation process in cells derived from healthy and diseased people, scientists could observe how that disease unfolds at a cellular level. They could also use the cells to test drugs that might correct those biochemical abnormalities. "We want to use these cells to ask and answer questions that can't be asked and answered any other way," says M. William Lensch, a research scientist at the Harvard Stem Cell Institute and Children's Hospital Boston.

The relative simplicity of the approach--and the fact that it can be supported by federal funding--means that many more scientists are likely to attempt reprogramming than cloning. (In 2001, President Bush limited federal funding for embryonic stem-cell research to embryonic stem-cell lines already in existence.) According to Story Landis, chair of the Stem Cell Task Force at the National Institutes of Health, in Bethesda, MD, the funding agency has already announced two programs to fund reprogramming research and would welcome applications to derive cell lines from patients.

While no one has yet announced that he or she has derived a disease-specific cell model, George Daley's lab at Harvard may be in the lead. Last month, he and his team published a paper in Nature showing that they can reprogram cells from a skin biopsy from a healthy person, and they are already trying to repeat the feat with tissue from patients. Ultimately, they are interested in developing models of sickle-cell anemia and Fanconi anemia, a hereditary disease in which the bone marrow doesn't produce enough new cells to replenish the blood.

For example, patients with Fanconi anemia often suffer from skeletal problems, and their cells show an impaired ability to repair DNA. "We don't have any idea why kids with DNA repair defect would get a blood disease, and why they sometimes get these bone abnormalities," says Lensch, who works with Daley. But with stem-cell lines developed from a patient, "we could push the cells to develop into bone and blood, and try to learn about the links between the two."

Such models could also help resolve long-held debates about specific diseases, such as Alzheimer's. By differentiating reprogrammed cells from Alzheimer's patients into neurons and comparing them with neurons derived from healthy embryonic stem cells or with cells with mutations that mimic a rare, hereditary form of the disease, scientists will be able to determine how much of Alzheimer's is due to the environment versus genes, as well as how similar the sporadic form of the disease is to the hereditary form. (Most drugs on the market for Alzheimer's were developed using models that mimic the hereditary form of the disease and have shown limited efficacy in patients.) "This is a whole new world of investigation," says Lawrence Goldstein, a neuroscientist at the University of California, San Diego, whose lab is about to begin collecting skin cells from Alzheimer's patients.

Despite the excitement, Lensch and others caution against abandoning other embryonic stem-cell research, especially therapeutic cloning. "We're in the early stages of this research, where we're excited about the possibilities but still need to show it's both useful and representative of the disease," says Snyder. In addition, he says, embryonic stem cells and perhaps cloned stem cells will be needed as controls for future studies.

Scientists also say that it's too soon to tell how easy it will be to generate stem-cell lines from patients: the genetic variations that lead to the disease could also impact the reprogramming process. "With some genetic disease, I think it will be really difficult," says Lensch.

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Gadis Australia Pasien Pertama Dunia yang Berubah Golongan Darah

Canberra (ANTARA News) - Seorang gadis remaja Australia --Demi-Lee Brennan-- menjadi pasien pertama yang mengubah golongan darahnya dan menerima sistem kekebalan dari donor organnya.

Brennan yang kini berusia 15 tahun menerima transplantasi organ hati pada saat usianya 9 tahun karena organ harinya tidak berfungsi.

"Hal itu adalah kesempatan kedua saya untuk dapat bertahan hidup," kata Brennan kepada media massa setempat ketika menceriterakan bagaimana tubuhnya berhasil menerima dan beradaptasi bedah transplantasi yang dapat dikatakan "Mukjizat" yang datang dari Tuhan. "Sungguh sulit dipercaya."

Golongan darah Brennan mengalami perubahan dari "O" negatif menjadi "O" positif pada saat ia sakit dan diberikan pengobatan untuk menghindari penolakan terhadap organ hati donor oleh sistem kekebalan tubuhnya.

Sel batang pembuluh darah hatinya yang baru memasuki sumsum tulang belakangnya yang mengubah seluruh sistem kekebalan tubuhnya, berarti si remaja Brennan tak lagi memerlukan obat-obatan anti penolakan tubuh.

Para dokter dari Rumah Sakit Anak Westmead di Sydney mengatakan mereka belum dapat memberikan keterangan kasus Brennan yang mengalami kesembuhan, seperti yang mereka sampaikan dalam majalah kedokteran, The New England Journal of Medicine.

"Terus terang kami belum menemukan penjelasan untuk hal itu," kata Michel Stormon seorang ahli hepatologi pediatri seperti dikutip Reuters.

Sturat Dorrney, mantan kepala bagian unit transplantasi di rumah sakit itu mengatakan, kasus Brennan dapat membuka jalan bagi terapi transplantasi organ, karena biasanya sistem kekebalan tubuh pasien penerima menyerang transplantasi jaringan di donor.

"Kini kami harus kembali mengkaji ulang semua tahapan yang terjadi pada Demi-Lee dan melihat mengapa hal itu dapat terjadi dan kalau-kalau dapat melakukan pengulangan kembali," kata Dorney.

"Kami berpikir hal itu mungkin karena kami menggunakan organ hati dari seseorang yang usianya masih muda dan Demi-Lee memiliki sel darah putih dalam jumlah rendah mungkin karena dua faktor itulahyang menjadi alasan," katanya kepada harian Daily Telegraph.

Penolakan tubuh umumnya ditangani dengan kombinasi obat-obatan walaupun penolakan kronik tidak terjadi dua arah (bolak-balik).

Hanya tujuh dalam 10 operasi transplantasi di Australia yang berhasil setelah lima tahun berselang yang dikarenakan oleh penolakan tubuh di pasien. (*)

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Pengobatan Kanker Juga Bantu Obati Osteoporosis

Washington (ANTARA News) - Obat yang digunakan untuk mengobati kanker tulang sumsum juga dapat membantu mengobati osteoporosis dengan merangsang sel-sel tungkai, kata beberapa peneliti AS, Jumat.

Mereka mendapati bahwa Velcade, yang dibuat oleh Millenium Pharmaceuticals Inc (MLNM.O) untuk mengobati tumor ganda di sumsum tulang, mengaktifkan sel-sel tungkai yang berubah menjadi tulang.

Ujicoba terhadap tikus memperlihatkan membantu mengaktifkan jaringan tulang dan mungkin merupakan pengobatan yang berpotensi bagi osteoporosis, kata satu tim di Massachusetts General Hospital dan Harvard Stem Cell Institute di "Journal of Clinical Investigation".

Ahli sel tungkai di Harvard Dr. David Scadden mengatakan, para ilmuwan telah berharap menemukan cara untuk menggunakan obat guna merangsang sel-sel tungkai, yang merupakan sel pengendali tubuh.

"Terapi sel tungkai seringkali dikira sebagai tindakan memasukkan sel baru ke dalam tubuh, tapi studi ini menunjukkan bahwa pengobatan dapat mengubah sel-sel tungkai yang ada yang terdapat di jaringan tubuh dan bertindak sebagai obat pengaktifan untuk meningkatkan mekanisme perbaikan sendiri tubuh," kata Scadden dalam suatu pernyataan.

"Obat yang mengarahkan sel-sel tak matang untuk menjadi sejenis sel khusus, seperti dalam studi ini, dapat berpotensi sangat bermanfaat," katanya seperti dikutip Reuters.

Velcade, yang secara generika dikenal sebagai "bortezamib", merangsang sel-sel tungkai "mesenchymal", demikian temuan para peneliti tersebut. Sel-sel itu berkembang menjadi zat pembangun-tulang "osteoblast" dan beberapa jenis sel lain termasuk "cartilage", lemak, kulit dan otot.

Ujicoba pada tikus memperlihatkan obat tersebut meningkatkan kegiatan "osteoblast", dan ketika digunakan pada tikus yang menderita osteoporosis, obat itu secara mencolok meningkatkan kepadatan dan susunan tulang.

"Jika paradigma yang terlihat dalam studi ini terbukti benar bagi jaringan lain, kita mungkin memiliki pilihan untuk memperbaiki dan mengaktifkan kembali berbagai tempat yang terpengaruh oleh cedera atau penyakit dengan menggunakan obat --itu akan sangat menggairahkan," kata Scadden. (*)

Copyright © 2008 ANTARA

Synthesizing a Genome from Scratch

By Emily Singer

In a technical tour de force, scientists at the J. Craig Venter Institute, in Rockville, MD, have synthesized the genome of the bacterium Mycoplasma genitalium entirely from scratch. The feat is a stepping stone in creating precisely engineered microbial machines capable of generating biofuels and performing other useful functions.

"It really is groundbreaking that you can synthetically build a genome for a bacterium," says Chris Voigt, a synthetic biologist at the University of California, San Francisco, who was not involved in the project. "It's bigger by orders of magnitude than what's been done before."

Biologists creating genetically engineered organisms now routinely order pieces of DNA that are 10,000 to 20,000 base pairs long--big enough to incorporate the genes for a single metabolic pathway. That allows researchers to engineer microbes that can perform specific tasks, but the ability to synthesize entire genomes could grant a whole new level of control over biological design. (See "Tumor-KillingBacteria.")

In the new study, scientists ordered 101 DNA fragments, encompassing the entire Mycoplasma genome, from commercial DNA synthesis companies. These fragments were designed so that each overlapped its neighboring sequence by a small amount; these overlapping stretches stick together, thanks to the chemical properties of DNA. Researchers then bound the fragments piece by piece, eventually generating the full 582,970 base pair Mycoplasma sequence. The findings were published Thursday in the online edition of Science.

"We consider this a second and significant step in a three-step process of our attempt to create the first synthetic organism," says Craig Venter, president of the Venter Institute. Venter and his colleagues ultimately want to create a minimal genome--one with the least number of genes needed to sustain life. Pinpointing the minimal genome will both shed light on key cellular processes and provide a base for designing sophisticated synthetic organisms. "We ultimately want to design cells that could function in a robust fashion to make unique biofuels," says Venter.

The researchers' next step will be to show that the synthetic genome functions as it should. "We have the whole genome assembled in a tube, but we need to transplant it into the cell of a different species to show that it can reboot the cell," says Hamilton Smith, a Nobel laureate who oversaw the project at the Venter Institute. Last year, Smith's group transplanted the genome of one species of Mycoplasma into another, demonstrating that this type of transplant is possible. (See "Transplanting a Genome.")

While the synthesis of a genome might be impressive from a scientific perspective, it is not yet a practical way to engineer microbes to make biofuels. Instead, several companies, including Synthetic Genomics, a biotech company founded by Venter to engineer microbes for energy, are using more traditional metabolic engineering techniques to generate fuel-producing bacteria. (See "Building Better Biofuels.") "What we're doing with synthetic chromosomes will be the design process for the future," says Venter.

Others in the field are excited about that prospect. "Being able to synthesize genomes opens up a new world," says Voigt. "You can build things on the scale of the genome." For example, he says, scientists are now engineering bacteria to perform different steps in the conversion of biomass into ethanol--one strain to break down the biomass, another to make ethanol. But ideally, scientists could put those processes together to create one organism that could eat biomass and spit out fuel. (See "The Price of Biofuels.") "That would require genome-scale design," Voigt says.

He likens the current project, which required multiple steps to glue the fragments together, to the last computers designed before automated manufacturing and microfabrication techniques were introduced. Similar advances are needed for more ambitious genome-synthesis projects. "We still need to develop 'one step' genome construction methods in order to reduce the costs and turn time of genome construction," says Drew Endy, a synthetic biologist at MIT.

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Treating Muscular Dystrophy with Stem Cells

Researchers at the University of Texas Southwestern Medical Center (UT Southwestern) have used embryonic stem cells from mice to grow muscle cells. These same cells, injected into mice with a mild form of muscular dystrophy, formed healthy, functional muscle fibers at the site of deteriorating tissue. Scientists say that the research, while still in its early stages, could eventually lead to a cell-based therapy for patients with muscular dystrophy and other muscle-related diseases. The research was recently published in the online edition of Nature Medicine.

According to the Muscular Dystrophy Association, about 250,000 people in the United States have some form of the disease. The most well known, Duchenne muscular dystrophy, is caused by a genetic mutation that disrupts the formation of dystrophin, an important protein involved in the formation of muscle cells. In the absence of dystrophin, muscles are unable to regenerate, and they gradually weaken and waste away. Eventually, the deteriorated area is taken over by fat and connective tissue.

Rita Perlingeiro, assistant professor of developmental biology at UT Southwestern, says that embryonic stem cells may be the key to reversing muscular dystrophy's debilitating effects. The advantage lies in the cells' pluripotency--the ability to transform into any mature cell, be it bone, muscle, or cartilage. However, many researchers have found it difficult to direct every stem cell in a culture to form a specific type of cell. In lab experiments, scientists often end up with a mixture of cells that, when injected into an animal, form large clusters resembling a tumor.

So Perlingeiro and her team set two main goals: to find the right set of cues to convert embryonic stem cells into muscle cells, and to look for ways to isolate muscle cells from the rest of the culture medium, in order to inject a dose of pure muscle cells into a mouse model.

In normal embryologic development, stem cells turn into various tissue and bone, depending on a combination of molecular and genetic signals. In the case of muscle cells, past research has shown that the gene Pax-3 is essential in pointing stem cells down the path of muscle formation. With this knowledge, Perlingeiro and her team grew mouse-derived embryonic stem cells in a culture dish, then genetically manipulated the solution to overexpress Pax-3. They found that, compared with mixtures without Pax-3, a significant number of stem cells exposed to the activated gene formed muscle cells.

However, not all of the cells turned into muscle, and when the team injected the solution into a mouse with a mild form of muscular dystrophy, the mixture caused tumors to form. The team then focused on developing an identification process that would make muscle cells stand out from the rest of the solution. Once again, Perlingeiro looked to basic developmental research and found that, during normal muscle formation in the embryo, stem cells that become very early versions of muscle cells display certain surface molecules, or markers. The team repeated the first phase of its experiment, exposing embryonic stem cells to Pax-3, and looked for the telltale markers indicating muscle cells. The researchers then isolated these cells, creating a solution that consisted solely of muscle cells.

In preparation for injecting the new solution into a mouse model, the team first injected cardiotoxin into the mouse's leg. The effect inhibited the production of dystrophin, causing a weakening of the muscle--a condition resembling muscular dystrophy. Perlingeiro and her colleagues then injected the mouse with the muscle-cell solution. The team then took muscle biopsies and, after immuno-staining, found that, compared with mice that did not receive the solution, treated mice exhibited more dystrophin, indicating healthy muscle regeneration.

To confirm their results, the researchers ran both groups of mice on a treadmill; they found that the mice that received the solution outlasted the group that did not. Perlingeiro went a step further: after sacrificing both animal groups, she and her colleagues extracted every leg muscle, treated or untreated. They then placed each muscle in a bath and tested its strength by exposing it to an electrical impulse. The team found that the stronger contractions came from the muscles treated with the stem-cell-derived solution.

Perlingeiro says that the study's results are encouraging, as she envisions one day providing stem-cell-based therapy for people with muscular dystrophy and other muscle-related diseases. However, there will have to be more follow-up studies before the technique can be applied to humans.

"I have a long to-do list," says Perlingeiro. "We'd like to use the same technique on human embryonic stem cells."

Recently, researchers were able to turn human skin cells into embryonic stem cells, a technique that bypasses the thorny issues currently surrounding use of embryonic stem cells. Perlingeiro says that combining this technique with her muscle-deriving method may one day yield effective, efficient treatment of diseases such as muscular dystrophy.

"If we can reprogram skin cells to become pluripotent, and use Pax-3 to make muscle, then we would be able to make cells from the patient, and we wouldn't face ethical issues or problems of rejection," says Perlingeiro.

Paul Muhlrad, a research program coordinator for the Muscular Dystrophy Association, says that the study's results are a promising step toward effective treatment for muscle-related diseases. "These researchers present a nice proof of principle that embryonic stem cells can be turned into muscle-producing cells in the laboratory and used to deliver healthy muscle to people with Duchenne muscular dystrophy," says Muhlrad. "Of course, these experiments were done with mice. We've yet to see whether they will work in humans, but this study offers us much hope."

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Mixing Mammals

By outfitting mice with a chunk of DNA that directs wing development in bats, scientists have created rodents with abnormally long forelimbs, mimicking one of the steps in the evolution of the bat wing. Their work gives weight to the idea that variations in how genes are controlled, and not just mutations in the coding regions of genes, are a driving force in evolution.

The slightly longer forelimbs of the transgenic mice "make them more batlike," says Nipam Patel, a professor of molecular and cell biology and integrative biology at the University of California, Berkeley, who was not involved in the work. "It seems like a subtle difference, but evolution works by these subtle differences."

The researchers focused on a gene, Prx1, that plays a part in the elongation of limb bones in mammals. The gene's expression is regulated by another sequence of DNA, called a Prx1 enhancer. To investigate how the enhancer shapes limb development, Richard Behringer, a professor of molecular genetics at the University of Texas MD Anderson Cancer Center, and his colleagues around the country put the bat version of the Prx1 enhancer into mice so that it controlled the mouse Prx1 gene. These transgenic animals developed forelimbs that were on average 6 percent longer than normal by the time they were born. It was a significant difference, although "the mice look like mice," Behringer says. "They're not going to fly out of the cage." The researchers report their work in the latest issue of Genes and Development.

To have any chance of flying, mice would have to develop very different forelimbs, like those of bats, which are longer and have membranes stretched between the bones. Behringer says that he'd like to try replacing the limb enhancers in mice with those from other animals, such as whales or wallabies.

Charles Darwin contemplated the evolution of different kinds of limbs in On the Origin of Species. Starting with a basic limb pattern, "successive slight modifications," he wrote, eventually produce the various mammal limbs we see today: human hands, bat wings, whale fins.

"We think what we've done is made one of those slight modifications," Behringer says. "Maybe during evolution you'd have a lot of those and the limb would get a lot longer, and maybe some of the tissue would be retained between digits, ultimately leading to the structures that would allow a bat to fly."

"It's a very nice demonstration of something that people have been suspecting now for some time: that regulatory sequences rather than changes in protein sequences sort of drive evolution," says Susan Mackem, who heads the Developmental Biology Unit at the National Cancer Institute's Center for Cancer Research. Mackem was not involved in Behringer's research.

Behringer's team also found something unexpected. When the researchers created mutant mice that lacked the mouse Prx1 enhancer, the animals developed forelegs of a normal length. That suggests that more than one enhancer controls the expression of the Prx-1 gene in mice, ensuring what Behringer calls a "regulatory redundancy."

"As long as there is one copy to do the work, the other copy can be creative," says Ann Burke, an associate professor of biology at Wesleyan University.

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How Important Is the Latest Cloning Feat?

Scientists at Stemagen, a small biotechnology company in La Jolla, CA, reported yesterday that they have for the first time generated cloned human blastocysts--early-stage embryos--from adult skin cells. This is the first step in generating stem cell lines matched to individuals, which are crucial for creating new cellular models of disease and potentially important for future tissue replacement therapies. (See "Next Steps for Stem Cells" and "The Real Stem Cell Hope".) The new findings also confirm that access to fresh eggs from healthy young donors is a key part of successful cloning. Lack of access to human eggs has been the major barrier in the field. (See "Human Therapeutic Cloning at a Standstill".)

Cloned blastocysts have been generated before, but from embryonic stem cells rather than from adult cells. Scientists theorize that embryonic stem cells are easier to turn into blastocysts because of their earlier developmental stage.

Experts in the field have had a mixed reaction to the new work. "It's a nice achievement, but in my view, they haven't crossed the bar," says Evan Snyder, director of the Stem Cells and Regenerative Medicine Program at the Burnham Institute in La Jolla. "The real test will be, can you generate cell lines that are stable and self-renewing and normal?" Others applaud the confirmation of the feasibility of human cloning. "The fact that it can be done is important," says Jeanne Loring, a stem cell scientist at the Scripps Research Institute in La Jolla. "It wipes away that blot on our scientific integrity," she says, referring to a massive fraud unveiled in 2005 in which South Korean scientist Woo Suk Hwang claimed to have generated stem cell lines from cloned human embryos. (See "Stem Cells Reborn".)

To clone an embryo, a process also called nuclear transfer, scientists first strip an egg of its genetic material. Then they insert DNA from an adult cell, such as a skin cell, into the egg. Through an unknown process, the egg turns back the clock on the adult DNA and begins to develop as a normally fertilized egg would. From the embryo, researchers could theoretically collect a specialized ball of cells that can be coaxed to turn into stem cells. So far, however, no one has successfully performed this feat.

Stemagen, a relatively unknown player in the field, probably owes its success to access to human eggs through a close association with a local fertility clinic. (The company was founded by a fertility specialist at the Reproductive Sciences Center in La Jolla.) "We were able to get access to high-quality oocytes and have them in the incubator within one to two hours," says Andrew French, Stemagen's chief scientific officer.

Egg donors and the intended parents gave eggs in excess of those needed for in vitro fertilization to the Stemagen scientists for research. Regulations in many states prohibit compensation for donated eggs for ethical reasons, a requirement that has slowed other cloning efforts.

Starting with 25 fresh oocytes, French and colleagues generated five blastocysts--five- to six-day-old embryos consisting of 30 to 70 cells. Rather than attempting to generate stem cell lines from the embryos, the researchers sent them to an independent company for genetic confirmation of their results. "They showed we had completely removed the DNA from the egg donor and replaced it with DNA from the skin-cell donor," says French. One blastocyst was confirmed as a clone via two DNA-fingerprinting methods, while genetic analysis of two others indicated the likelihood that they were clones.

The next crucial step will be generating stem cell lines from cloned embryos, which many stem cell scientists speculate will be the most challenging step. "That's likely where Hwang failed," says Synder.

French and colleagues are planning such experiments, with results potentially in the next eight to twelve months. "The quality of our blastocysts improved with each experiment," says French. Based on the success rate of previous attempts to make stem cells from regular embryos, he estimates that Stemagen will be able to generate a stem cell line from between five and ten cloned embryos and report the results in the next year. The company aims to sell or license the lines to pharmaceutical companies and others who would use them to test new drugs or develop new therapies.

While human therapeutic cloning has always been an ethically contentious area of research--partly because it requires the creation and destruction of human embryos--it has recently come under greater fire. After the announcement of new techniques for reprogramming adult cells so that they turn into stem cells without first forming embryos, some opponents called for a halt on embryonic-stem-cell research. (See "Stem Cells without the Embryos".)

However, researchers in the field emphasize the need to pursue all reprogramming techniques. "Even though there are other techniques to reprogram a cell that have gotten a lot of press, we still don't know how those compare with the reprogramming you actually see with nuclear transfer," says Snyder. "My feeling is, if we understand nuclear transfer better, we will be able to do the other kind of reprogramming more efficiently."

From here

Magnetic Cell Therapy

Stents are expandable stainless-steel scaffolds commonly used to prop open clogged arteries. But inserting a stent can damage an artery's inner lining, and stented arteries may reclose after several months, causing blood clots and possibly heart attacks. Now researchers at the Children's Hospital of Philadelphia have devised a way to use tiny iron-bearing nanoparticles and a magnetic field to direct cells with therapeutic properties to the sites of steel stents. The cells could help repair arterial damage and prevent clotting, among other things.

"Stents have been known to induce severe trauma," says Robert Levy, chair of pediatric cardiology at the Children's Hospital of Philadelphia. "Repairing blood vessels with cell therapy is a very important concept that can be realized with magnetic targeting."

Levy and his colleagues engineered nanoparticles, or tiny spheres, of polylactic acid, a biodegradable polymer used in sutures and other medical applications. The team then loaded each nanoparticle with a small dose of magnetically responsive iron oxide and inserted it into a bovine endothelial cell--a cell found in a blood vessel's inner lining. The bovine cells were genetically altered to express a fluorescent marker, making them easily detectable.

Next, the researchers surgically implanted small metal stents in the carotid arteries of live rats. They injected the rats with a solution of treated endothelial cells and created a steady magnetic field around each rat using two large, external electromagnetic coils. Levy says that the magnetic field he and his colleagues applied was less than a tenth of the strength of the fields generated by conventional MRI machines. After 48 hours, the team created images of the rat using bioluminescence imaging.

The researchers found that the magnetic field caused the cells to migrate to the metal stents under two scenarios: when cells were injected directly into the carotid artery, near the stent location, and when they were injected farther away, in the aortic arch, whence they could have branched out to all areas of the body. In tests that didn't use a magnetic field, the cells migrated throughout the body with little direction.

Magnetically directing cells, particularly endothelial cells, to the sites of metal stents may have a significant therapeutic effect, says Levy. During surgical implantation, stents tend to scrape off endothelial cells, whose normal functions include helping prevent blood clotting. Endothelial cells are also barriers to inflammatory cells. While inflammatory cells normally flock to an injury to help repair it, in the absence of endothelial cells, they build up excessively, creating arterial blockage. In recent years, stents have been engineered to release anticlotting drugs to prevent arteries from reclosing. But such drug-releasing stents have problems of their own, including preventing endothelial cells from regenerating.

"Two years ago, clinicians noticed that patients in significant numbers were having problems with these stents, probably because the endothelium wasn't properly healed," says Levy. "Clotting, myocardial infarctions, and sudden deaths occurred, and this has caused a big uproar over stent usage."

Levy hopes that magnetically directing new endothelial cells to blood vessels may solve many of the problems that stents currently face. His team plans to continue experimenting on rats, using endothelial cells derived from rats instead of cows, to minimize risk of rejection. Now that he has found a way to direct cells to metal stents, Levy is also looking at other potential therapies, including nitric oxide, which is known to relax and dilate blood vessels. He is currently engineering cells to genetically express enzymes that produce nitric oxide, and he will eventually load them with iron-oxide nanoparticles that will drive them to the sites of stents, further opening arteries.

Levy adds that the magnetic-based technique has applications outside of cardiovascular therapy. For example, in treating lung cancer, clinicians often use metal stents to keep airways open. However, a patient's tumor may continue to grow, eventually obstructing the passage despite the stenting. Magnetically targeted therapies could help deliver specific drugs to stent sites to treat tumors, in addition to keeping airways open.

"Metallic implants are also widely used in other areas, like orthopedics, for complex fractures, and correcting spinal curvature, where cell therapies could also be helpful," says Levy. "Steel implants are widely used in medicine, and there are all sorts of situations where applications could be used."

What's more, Levy envisions that such therapies can be applied using conventional MRI machines. The magnetic field generated by MRI cores is an order of magnitude more powerful than the ones Levy used in his experiments, so fewer iron-oxide nanoparticles could produce the same effect.

Robert Langer, Institute Professor at MIT, believes that Levy's technique is a promising step toward directed cell therapies. "They were able to localize more drugs into the targeted areas," he says. "I think it's a neat idea that has a lot of potential."

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