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Thread: Why the discovery of a method to grow rat embryonic stem cells is important

  1. #1

    Why the discovery of a method to grow rat embryonic stem cells is important

    http://www.physorg.com/news149345043.html
    Recipe for capturing authentic embryonic stem cells may apply to any mammal, study suggests
    December 24th, 2008 in General Science / Biology

    Researchers have what they think may be a basic recipe for capturing and maintaining indefinitely the most fundamental of embryonic stem cells from essentially any mammal, including cows, pigs and even humans. Two new studies reported in the December 26th issue of the journal Cell, a Cell Press publication, show that a cocktail first demonstrated to work in mice earlier this year, which includes inhibitory chemicals, also can be used to successfully isolate embryonic stem cells from rats.
    As the headline suggests, this method that was developed for the rat may well be applicable to all stem cells but the finding is important for other reasons that I will summarize below.

    1. Until this study, nobody was able to isolate and grow embryonic stem cells from rats. You would have thought that it was simple. After they have been growing mouse embryonic stem cells since the early 1980's, human embryonic stem cells since the late 1990's. In 1998, shortly after the mouse embryonic stem cells were grown, I had asked a colleague whether we could start doing rat embryonic stem cell research. He told me that while many groups have tried, noone had successfully isolated and cultivate rat embryonic stem cells. This was shocking since the rat is the second most common laboratory animal after the mouse. Why not? The answer to this question will come out as people start to investigate the new medium that Austin Smith and his group in Edinburgh used to grown the rat cells. By the way, the medium has a code name. The medium that had been discovered for human included the use of mouse feeder cells. Of course, several so-called defined culture media have now been devised for the human cells but they are still not satisfactory and most laboratories have been able to achieve at best a 25% success rate of getting embryonic stem cells out. Apparently, this medium allows a much higher success rate. So, it is possible that this medium will provide a breakthrough in terms of efficiency of growing embryonic stem cells from many species, rat and human included.

    2. Rats are the second most used laboratory species, after the mouse. Some 90% of spinal cord injury experiments are done with the rat and the rat model is much more developed than the mouse. The availability of rat embryonic stem cells will of course allow the first use of rat embryonic stem cells to treat rat spinal cord injury, instead of forcing investigators to use mouse or human, and therefore cyclosporin and immunosuppressive drugs. Much more important is that fact that having rat embryonic stem cells will now allow the cells to be genetically modified and then implanted into rat blastocysts to produce transgenic rats. This was a very difficult thing to do in the past and one could not do the things with rats that was possible in mouse. Almost all transgenic experiments were done with the mouse. Now, the experiments can be done in the rat.

    3. What allowed the success of this study was the development of a culture medium that prevents embryonic stem cells from differentiating. Embryonic stem cells are difficult to grow because they have one purpose in life and that is to differentiate into cells to produce an embryo. The cell culture medium must stop this without making the cell abnormal or strange in some way. The mechanisms that which the cells differentiate is at stake here. If this new culture media works on all species or more species, this may lead to the discovery of a universal anti-differentiating mechanism that can be applied to a whole variety of cells besides the rat embryonic stem cell. This may also allow large-scale culturing of the cells. When embryonic stem cells were first grown, people thought that the cells could be mass-produced. It turned out to be inordinately difficult to grow the cells. If you looked at them cross-eyed or treated them slightly differently, they would differentiate or die. Manufacturing the cells in large quantities would have been difficult.

    Wise.

  2. #2
    Hi Wise,

    This is a bit off-topic but I've never asked a researcher this question. How do you feel about working on live rats? I know you do all you can to reduce the pain for the rats but it's not possible to take it all away. I'm all for the use of animals in clinical trials but was just wondering how you personally felt about working on them. I know your work is for the greater good.

    I don't think you've ever written about this and maybe you'd like to. Here's an opportunity if you don't think it will hi-jack your thread too much.

    Oh, interesting thread... I didn't know there was any problem in isolating and growing rat stem cells. Good to hear they've created a cell culture medium that doesn't force them to differentiate or kill them. And that shows promise in other fields and regards.

    Bob.
    "Be kind, for everyone you meet is fighting a great battle." - Philo of Alexandria

  3. #3
    Interesting information and good news, Wise Young. Thanks!

    I could care less about the rats feelings. As far as I know, there's no shortage of rats in this world and if they can help a human recover from a SCI or other injury or disease, then let's test the hell out of them!

  4. #4
    Banned adi chicago's Avatar
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    test humans to find the answer for cures.
    sorry if i am wrong.
    • Dum spiro, spero.
      • Translation: "As long as I breathe, I hope."

  5. #5
    Quote Originally Posted by bob clark View Post
    Hi Wise,

    This is a bit off-topic but I've never asked a researcher this question. How do you feel about working on live rats? I know you do all you can to reduce the pain for the rats but it's not possible to take it all away. I'm all for the use of animals in clinical trials but was just wondering how you personally felt about working on them. I know your work is for the greater good.

    I don't think you've ever written about this and maybe you'd like to. Here's an opportunity if you don't think it will hi-jack your thread too much.

    Oh, interesting thread... I didn't know there was any problem in isolating and growing rat stem cells. Good to hear they've created a cell culture medium that doesn't force them to differentiate or kill them. And that shows promise in other fields and regards.

    Bob.
    Animal experiments are essential for both discovery and preclinical validation of therapies before clinical trials. Without the animal models, none of the therapies that we have now would have been discovered or found to be effective for spinal cord injury.

    To minimize the number of animals needed to be used for testing therapies, I have spent the better part of 20 years establishing and optimizing a rat spinal cord injury model that is precise and reliable, thereby reducing the number of animals that need to be done in order to show significant effects.

    The model and outcome measures are standardized around the world so that data can be shared. Finally, I have taught quarterly workshops for a dozen years, to train people how to do the model so that it produces reliable data.

    The model produces very precise and graded spinal cord injury with validated outcome measures, so 10 rats per treatment group can detect a 10% change in lesion volume and 12 rats are sufficient to detect a 1 point change in BBB scores at p<0.05.

    Regarding pain, the rats are anesthetized at the time of injury and thus are asleep when it happens. Experienced caregivers take very good care of the rats. The main problem, as you might imagine, is bladder. For the first few weeks after injury, the rats need to get their bladders checked and squeezed every day. The rats don't have feelings below the injury site.

    Some of the rats get what appears to be neuropathic pain. They bite the dermatomes just below in the injury level. As soon as we see this, we start treating the rats with daily oral 64 mg/kg of acetaminophen, which seems to work quite well in stopping the biting. If it does, we euthanize the animals.

    Wise.

  6. #6
    Banned adi chicago's Avatar
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    sir acetaminophen is tylenol ?i take daily ...helps me .
    will damage my liver but ease the pain.
    • Dum spiro, spero.
      • Translation: "As long as I breathe, I hope."

  7. #7
    Quote Originally Posted by adi chicago View Post
    sir acetaminophen is tylenol ?i take daily ...helps me .
    will damage my liver but ease the pain.
    Yes, it helps our rats. I have been thinking of giving it intrathecally (where it won't have any liver toxicity). Wise.

  8. #8
    Quote Originally Posted by adi chicago View Post
    test humans to find the answer for cures.
    sorry if i am wrong.
    adi chicago,
    we must realize that all life comes from the same source, so we should test on human's.

  9. #9
    Quote Originally Posted by Wise Young View Post
    Animal experiments are essential for both discovery and preclinical validation of therapies before clinical trials. Without the animal models, none of the therapies that we have now would have been discovered or found to be effective for spinal cord injury.

    To minimize the number of animals needed to be used for testing therapies, I have spent the better part of 20 years establishing and optimizing a rat spinal cord injury model that is precise and reliable, thereby reducing the number of animals that need to be done in order to show significant effects.

    The model and outcome measures are standardized around the world so that data can be shared. Finally, I have taught quarterly workshops for a dozen years, to train people how to do the model so that it produces reliable data.

    The model produces very precise and graded spinal cord injury with validated outcome measures, so 10 rats per treatment group can detect a 10% change in lesion volume and 12 rats are sufficient to detect a 1 point change in BBB scores at p<0.05.

    Regarding pain, the rats are anesthetized at the time of injury and thus are asleep when it happens. Experienced caregivers take very good care of the rats. The main problem, as you might imagine, is bladder. For the first few weeks after injury, the rats need to get their bladders checked and squeezed every day. The rats don't have feelings below the injury site.

    Some of the rats get what appears to be neuropathic pain. They bite the dermatomes just below in the injury level. As soon as we see this, we start treating the rats with daily oral 64 mg/kg of acetaminophen, which seems to work quite well in stopping the biting. If it does, we euthanize the animals.

    Wise.
    Hi Wise,

    Under the circumstances, it seems as though your research group is being as humane as possible to those cute and cuddly white lab rats. And very "frugal" in the way it generates and disseminates its rat data. Especially with the standardizing of the 'model and outcome measures' so the data can be shared around the world, minimizing duplication and the overlapping of test results.

    I'm sure you're aware of the way the cosmetics industry used to operate. Perhaps some still do. Using live animals helter-skelter, especially rabbits in its eye irritant testing, when other non-animal means were proven to be just as effective. Since they are very competitive profit driven entities, with corporate secrets to protect, the test results were never shared among the community so needless duplicative tests were performed on those poor helpless bunnies. And just so they could reap a bigger profit on their latest line of gaudy blue eyeliner or lash-lengthening mascara. Vanity products.

    It's amazing how accepting animals are of their injuries. A dog can get paralyzed and might in curiosity glance back at its hind quarters once or twice post-injury, then with a couple wheels strapped to it get on with its life. Seemingly unaware that anything is amiss. I wish it were that easy for human SCIs.

    Would it be possible to use the rats in a series of secondary tests? A twofer. Instead of squeezing their bladders why not catheterize them with eentsy-weentsy catheters and find a way to keep us SCIs UTI-free? We need an antiseptic that kills bacteria but doesn't harm mucosal tissue.

    Araitn, you're a cold dude!

    Good luck in your research Wise. And thanks.

    Bob.
    "Be kind, for everyone you meet is fighting a great battle." - Philo of Alexandria

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