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Category Archives: Stem Cell Experiments
Posted: June 3, 2015 at 10:40 pm
In 2006, Kazutoshi Takahashi and Shinya Yamanaka reprogrammed mice fibroblast cells, which can produce only other fibroblast cells, to become pluripotent stem cells, which have the capacity to produce many different types of cells. Takahashi and Yamanaka also experimented with human cell cultures in 2007. Each worked at Kyoto University in Kyoto, Japan. They called the pluripotent stem cells that they produced induced pluripotent stem cells (iPSCs) because they had induced the adult cells, called differentiated cells, to become pluripotent stem cells through genetic manipulation. Yamanaka received the Nobel Prize in Physiology or Medicine in 2012, along with John Gurdon, as their work showed scientists how to reprogram mature cells to become pluripotent. Takahashi and Yamanaka’s 2006 and 2007 experiments showed that scientists can prompt adult body cells to dedifferentiate, or lose specialized characteristics, and behave similarly to embryonic stem cells (ESCs).
Takahashi and Yamanaka worked together at Kyoto University. Takahashi was a post-doctoral researcher who had earned a graduate degree in biology at the Nara Institute of Science and Technology in Ikoma, Japan. Yamanaka had earned an MD from Kobe University in Kobe, Japan in 1987. In 2004, Yamanaka began working at Kyoto University as a professor, where he studied factors that help an organism fend off retroviruses, which are single-stranded RNA viruses that can incorporate their genetic material into the DNA of a host cell. Yamanaka and others hypothesized that retroviruses could influence somatic cells to become stem cells. Yamanaka worked to find new ways to acquire embryonic stem cells to avoid the social and ethical controversies surrounding the use of human embryos in stem cell research during the late twentieth and early twenty-first centuries. Yamanaka studied the work of John Gurdon, a researcher who had experimented with Xenopus frogs at the University of Oxford in Oxford, United Kingdom. Yamanaka claimed that Gurdon’s work in reprogramming mature cells in frogs (Xenopus) in 1962 influenced his own work in reprogramming differentiated cells.
Yamanaka also noted that experiments in cloning Dolly the sheep in 1996, conducted by Ian Wilmut, Angelica Schnieke, Jim McWhir, Alex Kind, and Keith Campbell at the Roslin Institute in Roslin, Scotland, influenced his work. The Dolly experiment showed that scientists could reprogram the nucleus of somatic cells by transferring the contents of the nucleus into oocytes that have had their nuclei removed, a technique called somatic cell nuclear transfer (SCNT). Other research groups such as Masako Tada’s group in Japan in 2001 and Chad Cowans group in Massachusetts in 2005 combined embryonic stem cells with somatic cells to produce pluripotent cells. After these experiments with somatic cells, Takahashi and Yamanaka hypothesized that there were common factors, genes in particular, which caused somatic cells to become pluripotent stem cells.
In 2006, Takahashi and Yamanaka selected twenty-four candidate genes as factors that they hypothesized could possibly induce somatic cells to become pluripotent, and they began to test them one at a time. They used retroviruses to insert each of the twenty-four genes into the chromosomes of differentiated mouse embryonic fibroblasts. Each gene was inserted near the mouse Fbx15 gene, a gene that embryonic stem cells express during development in mice. The newly inserted gene endowed mice with resistance to an antibiotic named G418. The researchers labeled the resulting retroviruses mixed with host DNA as retroviral factors. Takahashi and Yamanaka placed the retrovirus-infected cells into cell culture with G418 antibiotic and cells to provide nourishment, called feeder cells. If one of the infected cells showed G418 resistance, then the scientists would know that one of the twenty-four genes influenced the cell to become an embryonic stem cell-like cell. However, none of the cells showed a resistance to G418, so Takahashi and Yamanaka reworked their approach.
Next, Takahashi tried to insert into a fibroblast cell multiple retroviral factors instead of one at a time. The researchers added all of the twenty-four retroviral factors at the same time into mouse fibroblast cells. This time, there were twenty-two cell colonies that showed a resistance to G418, meaning that there were colonies in which the cells exhibited embryonic stem cell properties. After examination, Takahashi and Yamanaka concluded that the cells were similar to embryonic stem cells and duplicated themselves in similar periods of as embryonic stem cells. They named the cells iPS-MEF24, signifying pluripotent stem cells induced from mouse embryonic fibroblasts by twenty-four factors.
The next experiments aimed to identify specific factors responsible for the generation of iPS cells. To isolate these specific factors, the researchers removed retroviral factors one at a time from the original twenty-four, and each time they removed a factor, they repeated their cell colony procedures. If the researchers removed a factor and the resultant cell colony wasn’t resistant to antibiotics, they knew that the missing factor somehow influenced the generation of iPS cells. Takahashi and Yamanaka repeated their procedure until they found ten genes that, when combined together in cells, yielded colonies of cells with G418 resistance. They named those cells with the ten genes as iPS-MEF10 cells. Takahashi and Yamanaka found that of the ten genes, when they combined four genes in particular (Oct3/4, KIf4, Sox2, and c-Myc), they produced the most cells that were like embryonic stem cells. The scientists named the cells iPS-MEF4. Takahashi and Yamanaka deemed those four genes important in the role of iPS cell generation. They concluded that iPSCs are similar, but not identical to embryonic stem cells.
To determine how embryonic stem cells were different from iPSCs, Takahashi and Yamanaka used primers, or strands of nucleic acid that help to start the process of DNA synthesis, to promote replication of genes found in normal embryonic stem cells. If an iPSC had a normal embryonic stem cell gene, the primer would prompt the normal gene to replicate, and the scientists could then see that the iPSC had a normal gene.
Takahashi and Yamanaka continued their experiments and injected the iPSC samples into mice that had no body hair. These nude mice were a variation of the common mouse (Mus musculus), but they had an inhibited immune system and lacked the Fox1 gene. When the researchers injected iPSCs into the mice, teratomas, which are tumors with germ layer components, formed. The teratomas resulting from iPS-MEF4 injections differentiated into all three germ layers (ectoderm, endoderm, and mesoderm), including neural and muscular tissues, cartilage, and epithelium. These tissue types formed aggregates of pluripotent stem cells called embryoid bodies. From the teratomas, Takahashi and Yamanaka took some cell samples and cloned them. They inserted the cloned cells into blastocysts by microinjection and obtained four different embryos.
After analysis, Yamanaka and Takahashi found that the four embryos contained iPS cells that contributed to all three germ layers, providing further evidence that the four genes (Oct3/4, KIf4, Sox2, and c-Myc) helped produce cells that were the most like embryonic stem cells. Takahashi and Yamanaka observed that the iPS-MEF4 cells continued to be more similar to embryonic stem cells than to other iPS cells. After further experimentation, they concluded that the iPS cells they generated were pluripotent in mice, and therefore
provided the possibly of repeating a similar experiment in humans. Takashi and Yamanaka published the results of their experiment in 2006.
After their mouse experiments, in 2007 Takahashi and Yamanaka published the results of another experiment that detailed methods and results used to produce iPS cells with human cells. They used the same four genes from humans that were used in mice. Another group led by James Thomson at the University of Wisconsin in Madison, Wisconsin, published their findings on iPSC in humans. They found that four genesOct4, Sox2, NanoG, and Lin28were sufficient to reprogram human somatic cells into pluripotent stem cells. Independent confirmation of Takahashi and Yamanaka’s previous experiments with mice supported the hypothesis that scientists can generate and use induced pluripotent stem cells in a similar manner as embryonic stem cells. Scientists later used iPSCs in regenerative medicine to research treatments for various human diseases such as Parkinson’s disease, platelet deficiency, spinal cord injury, and macular degeneration.
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Induced Pluripotent Stem Cell Experiments by Kazutoshi …
Posted: April 30, 2014 at 12:50 pm
Teratomas – tumors that arise from what are essenitally partially differentiated stem cells
As South Dakota considers repealing the state ban on embryonic stem cell research, this issue is receiving more and more attention on a national level.
President Obama came to the White House last month carrying the campaign promise that he would reverse President Bushs 2001 ban on taxpayer funding of embryonic stem cell research (except for a few specific lines of embryonic stem cells already harvested).
The fact that Obama has not yet done so is a matter of consternation for those eager to destroy innocent human life in the hopes that stem cells derived from them might, maybe, someday cure various diseases in other humans. There may be some eagerness for those tax dollars and research grants, too.
Even as some work to put the taxpayers on the hook for this destruction of innocent human life and to lift state bans, adult stem cell research has for a number of years been producing successful treatments for various illnesses and injuries. These includemeningitis-relatedlimb damage, brain injury, stroke, retinaregeneration, heart tissueregeneration,angina,diabetes, bone cancer, nerve regeneration,cerebral palsy,cartilage regeneration, Parkinsons,kidney damage,liver cancer, lupus,multiple sclerosis,and leukemia.
Adult stem cell therapy also doesnt have the same treatment issues ESC does. It also doesnt involve the destruction of innocent human life, because the stem cells are taken from various areas of the persons own body.
Embryonic stem cell (ESC) research involves the harvesting of stem cells from human embryos. In the process, the human embryo is destroyed.
ESC also faces serious problems such as tissue rejection, the same tissue rejection seen in organ transplant recipients who must remain on ant-rejection drugs the rest of their lives to prevent their body from rejecting the foreign tissue.
ESC also has a problem with tumors. It seems ESC has a tendency to produce tumors in recipients of the embryonic stem cells.
An editorial at Investors Business Daily yesterday examines this problem.
Posted: March 14, 2014 at 3:45 pm
The green tips of these chromosomes are telomeres, whose length is a measure of cellular “aging” and determines how many times a cell can divide.
In a roomful of kids cancer specialists, like those listening to the keynote speech byGeorge Daley, closinganinternational pediatric oncology meeting in Boston, the Myc gene is better known as a mutated weapon of mass destruction.
But this driver of cancer growth is also part of a four-gene cocktail that can reprogram an adult skin cell back into an embryonic-like stem cell that holds great therapeutic potential.
Daleys team was among the first to report the creation of induced pluripotent stem cells (iPS cells) in human cells. All it takes is a skin biopsy or a simple blood draw to begin to realize the full potential for any patient, Daley said.
Bone marrow transplants from donors already save the lives of some kids with cancer. But Daley sees a future where genetic diseases can be cured by creating iPS cells from a childs own diseased cells, rendering them disease-free through genetic intervention, then growing them into healthy tissue that wont be rejected by the patients immune system. The goal is to create customized patient-specific stem cells, Daley said.
The road to transplantation therapy begins with understanding what diseased cells can be transformed into iPS cells, how transformation affects the cells, and how the genetic defects can be fixed. For now, iPS cells are tools for research into disease mechanisms and tools for drug discovery, Daley said.
Daley reviewed his groups work with several bone marrow failure syndromes in children that predispose them to cancer. The research has resulted in unexpected findings and prospects for regenerative medicine therapy.
In one effort, a team led by Suneet Agarwal, an attending physician in hematology at Childrens Hospital Boston, observed reprogrammed cells that seemed to cure themselves. They reprogrammed cells from a 3-year-old girl with a mitochrondrial disease known as Pearson marrow pancreas syndrome. In a lab dish, her iPS cells slowly segregated and, on their own, purged the mutant mitochrondrial DNA. From the purged cells, Agarwals team generated disease-free blood stem cells without any detectable mutant mitochondrial DNA. This biological phenomenon might explain how some patients outgrow their need for blood transfusions.
A second bone marrow failure syndrome, dyskeratosis congenita, features mutations in the gene for an enzyme that maintains telomeres, extensions of chromosomes that normally diminish with each cell division. The iPS cell reprogramming process works in part by activating this fountain-of-youth enzyme so Daleys team wondered if the disease defect would make cells resistant to reprogramming.
Surprisingly, it didnt. In fact, Daley reported, Agarwal found that the pluripotent state actually reversed the premature senescence of the diseased skin cells. The iPS cells steadily regrew their telomeres. The pluripotent state overrode the limiting genetic lesions, suggesting potential future therapy to replicate the enzymatic activity.
Originally posted here:
Stem cell experiments in genetic blood diseases Vector
Posted: February 7, 2014 at 7:47 am
By Sarah Boseley, The Guardian Thursday, February 6, 2014 19:09 EST
Researchers say they have reversed equivalent of type 1 diabetes in mice using stem cell transplants
Scientists believe they may have moved a step closer to a cure for the type of diabetes that develops in childhood and usually leads to a lifetime of insulin injections.
Researchers in California report that they have reversed the equivalent of type 1 diabetes in mice through transplants of stem cells. Their experiments have replaced cells in the pancreas damaged by the disease that are unable to make insulin.
Without insulin, the body has difficulty absorbing sugars such as glucose from the blood. The disease usually first shows in childhood or early adulthood and used to be a killer, but glucose levels can now be monitored and regulated with insulin injections.
Scientists have long wanted to try to replace the damaged -cells that normally produce insulin. This has been one of the prime targets of stem cell experiments. But until now, it has proved difficult, partly because mature -cells do not readily regenerate.
Writing in the journal Cell Stem Cell, scientists at the Gladstone Institutes in San Francisco describe how they took a step back and collected skin cells, called fibroblasts, from laboratory mice. Then, by treating the fibroblasts with a unique cocktail of molecules and reprogramming factors, they transformed the cells into endoderm-like cells. Endoderm cells are a type of cell found in the early embryo, and which eventually mature into the bodys major organs including the pancreas. Using another chemical cocktail, we then transformed these endoderm-like cells into cells that mimicked early pancreas-like cells, which we called PPLCs, said the Gladstone postdoctoral scholar Ke Li, the papers lead author. Our initial goal was to see whether we could coax these PPLCs to mature into cells that, like -cells, respond to the correct chemical signals and most importantly secrete insulin. And our initial experiments, performed in a petri dish, revealed that they did.
The team then injected these cells into mice that had been genetically modified to have high glucose levels, mimicking the type 1 diabetes condition in humans.
Importantly, just one week post-transplant, the animals glucose levels started to decrease, gradually approaching normal levels, said Li. And when we removed the transplanted cells, we saw an immediate glucose spike, revealing a direct link between the transplantation of the PPLCs and reduced hyperglycemia [high glucose level].
Scientists move closer to stem cell cure for Type 1 …
Posted: November 27, 2013 at 1:43 pm
10.17.2013 Joshua Hare Wins Grant to Take Stem Cell Experiments to Space
Through his groundbreaking research that showed stem cell therapies repair damaged hearts, the Miller Schools Chief Science Officer Joshua Hare, M.D., already shattered the earthly view that heart muscle cannot rejuvenate. Now, armed with a new grant from the organization that manages research aboard the International Space Station, Hare is ready to expand his stem cell therapy research to the final frontier outer space.
Hare, the founding director of the Interdisciplinary Stem Cell Institute and the Louis Lemberg Professor of Medicine, is among seven stem cell researchers across the nation who this week were awarded up to $300,000 each for the opportunity to use the space stations unique environment to explore the impact of microgravity on fundamental stem cell properties.
We believe that microgravity could play an important role in generating new heart muscle, Hare said. We are thrilled that this grant gives us the opportunity to test that theory.
Until NASA certifies Hares CASIS proposal, The Effects of Modeled Microgravity on c-kit+ Cardiac Stem Cell Division and Differentiation, as flight-capable, he and his team will conduct ground-based experiments comparing c-kit+ cardiac stem cells cultured in normal gravity to the same cells grown in a three-dimensional rotary cell culture system, which simulates microgravity. Part of the grant will be used to purchase the 3-D system.
The investigators assume that a 3-D microgravity environment will enhance the survival, migration, proliferation, and commitment of c-kit+ cardiac stem cells, which are identified and isolated by their c-kit+ marker, to myocardial lineages, giving them a greater ability to turn into heart muscle and reverse damage from heart attack and heart failure.
In our work, we have discovered that mechanical forces and 3-D architecture play important roles in stem cell biology and, for these reasons, we hypothesize that the three-dimensional microgravity environment will enhance the survival and proliferation of cardiac stem cells, offering opportunities not only to enhance production of cardiac stem cells for therapeutic purposes, but to understand the medical implications for individuals spending prolonged periods in microgravity, Hare said.
A pioneer of stem cell therapies for heart attack and heart failure, Hare led the seminal studies that showed that injecting bone-marrow derived mesenchymal stem cells (MSCs) from either the heart patient or a donor during bypass surgery could do what cardiologists were long taught was impossible: repair damaged hearts, and restore heart function.
In a more recent paper, published in the journal Circulation, Hare and his team demonstrated in a large animal model that injecting a mixture of MSCs and cardiac stem cells (CSCs) into the area surrounding an infarct reduced scarring twice as much as using MSCs or CSCs alone. The combination, which also led to an even greater increase of restored heart function, will soon be slated for clinical trials in humans.
Now Hare and his team hope the unique microgravity environment of the International Space Station will shed new light on how mechanical forces influence the generation of new heart muscle from CSCs, leading to the development of novel, less invasive, and less expensive stem cell therapies for people with heart failure.
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Joshua Hare Wins Grant to Take Stem Cell Experiments to Space …
Posted: November 26, 2013 at 10:41 pm
healthsection Health Topics A-Z Healthy Living Featured Conditions eHow Now eHow Health Conditions & Treatments Brain & Nervous System Stem Cell Experiments
Garrett Daun started writing professionally in 1993. Daun has extensive training in meditation, rock climbing, yoga, martial arts, exercise and massage therapy. His work has appeared in “The Squealor,” the “Earth First! Journal” and on numerous websites. Daun earned a Bachelor of Arts in religious studies and creative writing from the University of Oregon. He is a yoga and Radical Undoing trainer.
On March 9, 2009, the Bush Administration policy restricting federal funding for human embryonic stem cell research was overturned by Barack Obama in the name of easing human suffering. Stem cell research using induced pluripotent stems cells cloned from cells obtained from adult skin and donated embryonic stem cells continues to gain funding and attention from major researchers and funding foundations around the world. Stem cells have the ability to become tissue cells in various different organs of the body. Under certain conditions, stem cells can be induced to repair and replace damaged cells and tissues in various diseases and injuries.
Embryonic stem cells were first isolated in mice by researchers at the University of California, San Francisco and the University of Cambridge in 1981. Researchers at the University of Wisconsin and Johns Hopkins University isolated human stem cells from donated embryos 17 years later in 1998. In early 2001, President Bush put a hold on federal funding of all stem cell research. By the middle of 2001, federal funding was restricted to stem cells cloned from existing lines. Bush vetoed bills that would have expanded federal stem cell research funding in 2006 and 2007. Near the end of 2007, scientists at Kyoto University in Japan and the University of Wisconsin, Madison, published studies demonstrating a method of obtaining stem cells from adult skin.
A majority of funding for stem cell research comes from federal and state organizations like the National Institutes of Health and nonprofits including the Christopher and Dana Reeve Foundation. Many state governments have taken responsibility for funding stem cell research at state universities, institutions and businesses. A California ballot measure supported by Governor Schwartzenegger provides $300 million per year for 10 years for stem cell research and experimentation and Connecticut provides $10 million worth of annual funds toward stem cell research for curing disease. In 2004, New Jersey became the first state to support stem cell research by giving $9.5 million toward the development of the Stem Cell Institute of New Jersey. Maryland, Missouri and Iowa are among the states who pioneered stem cell research funding programs in the early to mid 2000s.
Hans Keirstead, Ph. D. is one of the world’s leading stem cell researchers who conducts animal and human stem cell research in an attempt to reverse the effects of spinal cord injury and paralysis. Raymond D. Lund at the University of Utah and Robert Lanza of Advanced Cell Technology in Massachusetts developed a repeatable method of inducing embryonic stem cells to become retinal pigment epithelium cells for treatment of macular degeneration and similar eyesight impairing diseases. A team of researchers at the Boston Children’s Hospital and Harvard Stem Cell Institute discovered that the process of creating induced pluripotent stem cells has the side effect of restoring and improving the health of the cells.
Proposed and experimental stem cell treatments include reversal of aging, spinal cord injury damage and brain damage. Research is underway to develop cell-based treatments for heart disease, diabetes, lung ailments, skin disorders, bone marrow deficiencies, blood disease, nervous system disorders, and organ repair and restoration. Most research involves exploration of treatments in animal models, although as of June 2010, Hans Keirstead of UC Irvine is conducting small scale human trials of a treatment method for recent spinal cord injury patients and has future plans for testing treatments for spinal cord injuries sustained years earlier.
While Hans Keirstead and other leading stem cell researchers warn that stem cell-based cures for paralysis and brain injuries might still be decades away, the potential for such cures is becoming increasingly anticipated by researchers and patients alike. Human stem cells have the potential to rejuvenate cells of every organ and tissue in the body if scientists can create the proper conditions for their use, which could supplant the need for organ transplants. Stem cells are increasingly used to test new medications for safety and effectiveness. The most far-reaching stem cell hope is the possibility of reversing aging to attain something close to cellular immortality.
The World Health Organization reports that 12.2 percent of the worlds people die from respiratory and lung diseases. Many more are partially…
Posted: November 20, 2012 at 1:46 am
The views expressed in this column do not necessarily reflect the personal views of the reporter, the Blue Print newspaper or Moffat County School District. Reporters are asked to take a specific position in order to share selected perspectives.
NO – by Bear Steadman
When Albert Einstein discovered how to split an atom, he envisioned scientific breakthroughs that would better mankinds knowledge of life and the universe. Instead, his research was used to create the atom bomb, which is capable of only one thing, causing death and destruction. So whos to say a similar outcome wont come from stem cell research? It doesnt matter how good someones vision or intentions are, we cannot ignore the fact that this research could be used for the wrong reasons.
A large percentage of older scientists are against furthering progress in stem cell research because of the potential negative effects. Many feel messing with human stem cells and DNA can only have detrimental results. There is proof that in some stem cell experiments, the participants suffer horrible side effects. For example, since 1981, the use of embryonic stem cells has had a common result, the development of tumors in the area the stem cells were injected. Because of this, many scientists and government officials feel that it is necessary that we discontinue funding and research in this field.
We also have to think about the possible side effects of genetic engineering on the human race. It is a well known fact that DNA changes over time. If everyone were to start getting genetically engineered body parts that could possibly contain insignificant genetic differences, this could cause huge mutations in our DNA in future generations. If the DNA in a genetically engineered ear were slightly altered, wouldnt that leave room for the possibility of genetic mutations for that person or maybe even their kids some day?
Another extremely controversial issue involving stem cell research is animal rights. Experimenting on animals has been a big issue for quite a while and will become an even bigger if we continue to experiment with stem cells. Scientist are now able to grow human body parts on animals. So far they have only been able to replicate small body parts. The reason behind this is to see if the rats body would support and grow a foreign body part, but obviously at the animals expense. Although there will never be a rat to human transplant (the ear would not survive after being detached from the rat) growing human body parts on animals is cruel and unfair. The goal of science should be to better our understanding of life and the universe, to see how things work and how we can use them to make the world a better place, not to torture animals with cruel experiments or to genetically manipulate life.
We need to ask ourselves these kinds of questions before we jump into research that can have such a major negative impact. One small mistake or miscalculation could lead to one big catastrophe and we possibly could face some serious problems in our future. This decision is ultimately up to our generation.
YES – by Sarah Dippel
Over the years, science fiction movies have made fun of cloning super humans. They were stories purely for the amusement of the audience. In the late 1960s however, stories that people thought were merely fictional started coming true.
Stem cell research has opened many doors that have been locked for the past 50 years. The cure for diseases, preventing birth defects, growing new skin, and even new appendages for burn victims and amputees had scarcely been thought of until stem cell technology developed. The recent research scientists have been performing is giving them the opportunity to do all of these things. Skin as well as organs such as noses and ears would be available for burn victims.
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Is stem cell research ethical?
Posted: March 13, 2011 at 8:34 pm
michaelsavage.com Savage on Obama’s executive order giving federal funding to embryonic stem cell experiments which much of the citizens of the USA oppose. And for good reason.
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Savage on Embryonic Stem Cells and Obama