Scientists Can Grow Stem Cells In a Petri Dish

rift321 writes "Scientists safely created induced pluripotent stem cells from human stem cells, and grew them in a petri dish. The previous methods for creating iPSC's involved the use of retroviruses, which rendered the stem cells unacceptable for human implantation due to an increased risk of cancer and mutations. The researchers used a safer, albeit slower process to modify the skin cells, using a cell-penetrating peptide to deliver the needed genes into the cell (PDF). I'd like to hear if anyone has some insight into exactly how close that brings us to everyday-use of stem cells for regenerative therapy, and exactly what obstacles remain before such therapies can be put to use."

Adult stem cells is the answer.

[reporter]

This breakthrough is important because ultimately adult stem cells is the answer.

Though fetal stem cells (taken from aborted fetuses) may be useful for research, organs grown from them cause severe rejection in the recipient of the transplant.

The only way to overcome this rejection is to grow the organs from the adult stem cells taken from the recipient herself.

Re:Obstacles?? OBSTACLES???

[ColdWetDog]

There's only one real obstacle. Make it affordable.

And just what do you think you're going to do with it? TFA is a stupid, hype filled mess (as usual). The original article is here [cell.com]. The big deal is thus:

- Pluripotent stem cells MAY (big time maybe) be useful in a wide array of clinical applications.
- Embryonic stems cells are hard to obtain, have ethical issues to some folks.
- Fibroblasts (skin cells) are easy to obtain, not so ethically high strung and easy to grow.
- We've known for a couple of years that you can create what look to be stem cells by infecting fibroblasts using four (count'em only four) separate proteins. That's pretty amazing - the whole differentiation cascade is apparently controlled by a small number of discrete molecules.
- The problem is that to get those proteins inside the fibroblasts you have to use a (real) virus, which is interesting called a "Trojan" since it's "bringing in" foreigners. That is conceptually unappealing because the "stem cells" now have some viral garbage which may or may not interfere with experiments.
- The current research gets said proteins inside the cell using another clever "hack" - a small peptide (apparently derived from HIV infected cells) that allows the bigger proteins to sneak inside and do their magic.

- On first and second blush, the newly transformed stem cells look and act like Pleuripotent Embryonic Stem Cells (the real McCoy).
- But this remains to be seen, many a grant and paper are yet to come.

So don't get all wound up thinking you can inject this stuff into you and magically turn 16 again. Remember, you can only be young once.

But you can be immature forever.

Depends

[ceoyoyo]

how close that brings us to everyday-use of stem cells for regenerative therapy, and exactly what obstacles remain before such therapies can be put to use.

It depends what you mean. Stem cells are being used for a variety of therapies at the moment. If you mean growing complete organs then you probably have a while to wait. It's one thing to grow some cells in a petrie dish and quite another to convince them to organize into, say, a heart.

Of course stem cells can be grown in a petri dish!

[Anonymous Coward]

The innovation here is that they have a new approach to transform the cells into stem cells that may be safer than previous alternatives. For example, gene therapy commonly relies on viral vectors to insert genes to produce the proteins into the genome. However, because these insert randomly, they can inactivate genes involved in cell proliferation regulation etc. resulting in cancer. There are other approaches such as naked DNA transformation, but then the genes producing the proteins are generally not replicated or segregated evenly when the cell divides and are thus lost over successive divisions.

What these people have done, is to avoid all the usual problems by making the required proteins (already well known) for cell transformation in a bacterial system and adding a seqeuence to them that produces a cell penetrating end - similiar to that found in some viral proteins. This allows their proteins to penetrate the cells and activate pathways that inactivate/activate certain genes sets to make the cells pluripotent. These changes appear to be permanent and hold for over 35 passages.

As a side note, this is a burgeoning field of research. The efficiency and efficacy of certain protein products as above, and even genetic material, can be greatly enhanced by the addition of nuclear localisation sequences, certain histones and so on, without nasty side effects.

Stem cells are not a cure

[Anonymous Coward]

Stem cells are a treatment technology, or may lead to one. One of the likely targets for therapy is to replace lost cells in the substancia negra in the brain that occurs in Parkinson's disease. Having the stem cells means you have to induce them to develop into s-n types cells, find a way to put them into the center of the brain, avoid immune responses if any and infection from the operation. It would also be nice to see the new cells did not become malignant or die quickly due to the same things that caused the s-n cells to die in the first place. A researcher on a panel discussion part of the NYC PD Unity Day events guessed less than 10 years.

Re:Of course stem cells can be grown in a petri di

[Anonymous Coward]

The factors are not made in a bacterial system BTW. They are expressed in HEK293 (derived from human embryonic kidney) cells and the cell extract used.

Breakthrough yet to be optimized

[modrzej]

The whole idea is pretty simple: just delivering four key reprogramming proteins using shuttle of cell-penetrating peptide. Basing on experience of everyday life we may suppose that a simple solution is free of interference from large number of unknown factors, thus efficient. But that's not the case, the protocol developed by the authors leads to transformation of mere 0.001% of input cells, which is order of magnitude less than in protocols based on viral transfection, and perhaps orders of magnitude less than threshold for applications in medicine. Some improvement could be gained, however, if purified proteins were used. Moreover, this fibroblasts were used to some extent as "blackboxes" with transformation-inducing proteins provided and results checked out, but with no developed sense of what's going on inside, which constitute room another room for improvement.

Clever idea

[pesho]

The idea is very clever. The technique itself has been around for quite some time, but nobody has tried to use it to reprogram cells into iPSC. The main advantage is that you eliminate introduction of viral DNA/RNA into the cells, which has been causing problems. The disadvantage is that it is significantly less efficient when compared to viral vectors. I don't think this is a problem.There is a lot of room for optimization.

There is more to this than it is published in the paper. You can use the same trick to push your newly obtained iPSC to differentiate into the cell type you need. For example introduction of MyoD can turn them into muscle cells for treatment of muscular dystrophy.

Everything Gives You Cancer

[Yergle143]

Based on the words of my stem cell buddies, making stem cells is relatively easy. The hard part is differentiating them into the tissue that you want -- safely. See if you inject stem cells into an animal (or a person http://www.the-scientist.com/blog/display/55430/ [the-scientist.com]) you generally get a tumor. This creates a new paradigm of medicine. To get approved a normal drug goes through three phases of evaluation where phase I is "safety". With Stem cell treatment, Phase I is a very big deal. 537

Re:Real response to troll

[Insanity Defense]

you are aware that embryonic research has not been banned in the US, just no more tax dollars, right?

you are aware that embryonic research has not been banned in the US, just no more tax dollars, right?

You are aware that it went a little further than that aren't you? The block was extended to using anything that had ever had Federal money used on it. So if you used part of a prior grant to equip your lab for different research you had to scrap it all and start again. You couldn't even buy used equipment (or buildings) that might once have been paid for by Federal money.

Since most research facilities that could do this work had at some time used Federal money to do the research meant building a new lab and equipping it with all new equipment. That drove the "buy in" cost of doing the research way up.

Of moderate importance

[Mutatis Mutandis]

Creating stem cells from adult cells is so far mainly interesting for research purposes. The first hope of researchers is that in the petri dish, culture flask or microtiterplate, stem cells and stem cell derived cell lines may be better research tools than the current cell lines. The cell lines currently used in laboratories are often cancer cell lines and poorly representative of the cells in someones brain or liver. Stem cells may also help us to better understand cell development and what happens when it goes awry.

For therapeutic applications, the first applications may depend on finding drugs that stimulate stem cells to differentiate: It may not be necessary to inject stem cells or cell derived from stem cells, because we may all carry cells with a differentiation potential. For example, regions of the brain seem to contain cells that could potentially differentiate to help people who are suffering brain damage or degeneration.

However, controlling differentiation is complex. While only four factors are sufficient to induce any cell to revert to a stem-cell state, inducing a stem cell to become e.g. a motor neuron is a very complex process that needs to be controlled step by step.

Re:Wow...

[interkin3tic]

One potential problem is that they're using c-myc, an oncogene. [wikipedia.org]

From the actual paper

Here we report the generation of stable iPSCs from human fibroblasts by directly delivering four reprogramming proteins (Oct4, Sox2, Klf4, and c-Myc) fused with a cell-penetrating peptide (CPP).

Tumor cells are often found to have more c-myc protein than they should, and mice which have too much c-myc develop cancer much more frequently. By getting protein in rather than genes expressing the protein into cells, this is probably safer, as the cells aren't going to continue making c-myc presumably, but it is something that needs to be rigorously tested.

I think the chances of it being anything really dangerous are pretty low. You're going to be turning individual cells with this, growing a whole bunch, and differentiating the cells before putting it into a patient. The protein only needs to work on that first cell, and proteins are turned over pretty quickly, it's likely that by the time you got to making new tissues it would already be gone, and any left over would probably be very dilluted to where it won't have an effect.

Also, you can make these cells without it. C-myc was one of the initial four genes used to make plurpotent cells, but a short while later it was shown that c-myc only increases the efficiency. This current method is low-efficiency, and you'd expect the efficiency to drop even lower without c-myc. As the summary points out, they're already working on ways to increase the efficiency, dollars to doughnuts they're going to be trying it without c-myc in the near future.

As far as the question in the summary

I'd like to hear if anyone has some insight into exactly how close that brings us to everyday-use of stem cells for regenerative therapy, and exactly what obstacles remain before such therapies can be put to use."

It puts it a lot closer. Transgenic tissues are much more worrisome than protein-treated tissues. When you give the cell new instructions for how to make proteins that can cause cancer, that's dangerous. Supplying those proteins yourself, not as much.

There is still one major obstacle that's probably a bigger concern: we can deprogram these cells, but you want to be absolutely sure though they're reprogrammed before you put them into a patient. If you want to put new neurons into someone, you have to be sure you've turned all the iPSC into neurons. If you put undifferentiated cells into someone's brain, they tend to develop teratomas, which I think is one of the worst types of tumors to have. They're making strides on that, but I don't think they're to the point where they can say for sure that's not going to happen.

This current breakthrough is absolutely a great thing though, don't mean to diminish it, just that there are other steps.

Re:Wow...

[interkin3tic]

I would have to disagree with a Teratoma being the worst type of tumor to have (albiet in ones brain of course it could present some serious issues) considering they are mostly benign and very treatable. And on the cool factor you can find some rather interesting things inside of them. I should also say gross factor, as most people do not share my ideas of what is 'cool'

You're talking about the teeth and hair? Yeah, that's gross and cool in my book.

I would wonder how long it would be until we can be absolutely sure that we have successfully reprogrammed these cells. I'm not sure that is possible at this point, though someone feel free to correct me on that, as I am not sure of the process.

Yes, it seems like it's the same as stem cell differentiation after the deprogramming, and we've been working on that for some time. You can definitely get stem cells and IPSC to differentiate in culture prior to implanting. For instance the yamanaka paper or the other initial paper detailing iPSC showed you could direct differentiation into cardiac like cells which would beat in the dish. And they had other ways of showing they directed differentiation of some tissues.

However, making a pure culture is an issue for at least neurons. Not really a good way (as of the last review paper on neurogenesis I read, a year old so I could easily be out of date) to make sure you've gotten all the cells to become postmitotic neurons. I know you can get cultures that are enriched with neurons and are at least mostly neurons, but AFAIK implanting that into a spinal cord lesion might get you teratomas there because of a few cells we can't isolate out yet. And there are probably issues with organization of neurons after that as well.

I realize that I'm thinking only of neurons, since I work on neurogensis (sorta). I should have said that I don't think we're there for brain and spinal cord, but don't know about other tissues. Maybe we've figured out how to turn stem cells into all liver cells already. Maybe pancreatic cells, I think I remember hearing something about that. Anyway, I'd amend my statement to "we might be ready for clinical trials with some non-neural tissues with this finding."