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Cancer Stem Cells??


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#1 ChaChi

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Posted 27 July 2010 - 02:40 PM

Hello,

I'm a high school student, and I'm extremely interested in doing a cellular biology project for science fair this year. Last year I did one using MCF-7 cells and kombucha tea. This year I want to do something much more advanced. I've been looking into stuff with Cancer Stem Cells, but I'm confused. Can you work with these cells in most labs? or is extensive equipment needed? What exactly is involved in Cancer Stem Cells?

I had been reading some abstracts involved with it, and it sounded realitively normal, but I'm wondering if I may have misunderstood.

If this is something that is completely out of my league, does anyone have any recommendations as to what I could do? Please keep in mind that I'm trying to push myself as much as possible, not looking for something simple. I participated in INTEL ISEF last year, and I found that students were able to pull off things adults thought impossible.

Thanks,
ChaChi

#2 bob1

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Posted 27 July 2010 - 04:25 PM

Cancer stem cells are the "precursors" of cancers in a fashion, sort of the seeding population that actively divides and can migrate. They are dificult to obtain and maintain, though not as much as embryonic stem cells. The techniques used for handling are pretty similar to ordinary culture practices, though there are things like feeder cells and the need to extensively characterise to ensure that they have not lost the stem cell phenotype.

To get them you need a fresh cancer mass (getting hold of fresh tissue is always an issue) of some sort, and processes for getting the stem cells out. You would need some experience in doing this and growing the cells in the appropriate medium, which I guess means that you would need to be in a lab for most of the proceedures. There are several lines out there that will be more or less stable as cancer stem cells, which it may be possible to work with. Most stem cells have a finite life span.

Something that is probably more achievable is to grow primary fibroblasts either as embryonic fibroblasts or skin fibroblasts, and then do some experiments on them. You may also be able to do something with induced pleuripotent stem (iPS) cells, as these are relatively easy to make and maintain.

#3 ChaChi

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Posted 27 July 2010 - 06:03 PM

Thank you, that response was extremely helpful. The professor with whom I usually work is on vacation and I'm doing my own research. I'll look more into what you told me.

Does anyone have any examples of experiments done with these cells? I'm not looking to copy anything, I just want to get ideas. I was so lucky to stumble upon this site while doing research. This forum has been more helpful than many of the sources I've been looking at.

#4 Piersgb

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Posted 28 July 2010 - 04:07 AM

Bob1, you mention iPSCs. What experience, if any, do you have with these? I'm currently on a project looking at culturing Mouse Bone Marrow Cells and to expand the population of hematopoietic stem cells to then be able to treat them with IL-5 to generate an eosinophilic-like population of cells. I've had quite a lot of issues with getting the culture to work well, so maybe looking at/using iPSCs could be an option. What do you think?

#5 Rsm

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Posted 28 July 2010 - 06:11 AM

Cancer stem cells are the "precursors" of cancers in a fashion, sort of the seeding population that actively divides and can migrate. They are dificult to obtain and maintain, though not as much as embryonic stem cells. The techniques used for handling are pretty similar to ordinary culture practices, though there are things like feeder cells and the need to extensively characterise to ensure that they have not lost the stem cell phenotype.

To get them you need a fresh cancer mass (getting hold of fresh tissue is always an issue) of some sort, and processes for getting the stem cells out. You would need some experience in doing this and growing the cells in the appropriate medium, which I guess means that you would need to be in a lab for most of the proceedures. There are several lines out there that will be more or less stable as cancer stem cells, which it may be possible to work with. Most stem cells have a finite life span.

Something that is probably more achievable is to grow primary fibroblasts either as embryonic fibroblasts or skin fibroblasts, and then do some experiments on them. You may also be able to do something with induced pleuripotent stem (iPS) cells, as these are relatively easy to make and maintain.


I have never came across a feeder cell co-culture of cancer stem cells... Most people (at least in melanoma research) try to immediately inject them into mouse. And then it depends on the mouse, how "good" your CSC are. See for example http://www.ncbi.nlm....les/PMC2597380/ (Quintana 2008), hotly debated. I think most people agree that cell lines do not contain CSC. So you'd need access to fresh tumour samples, a FACS sorter and a severely immunocompromised mouse. I guess that'll be difficult.

Cheers,

Minna
I got soul, but I'm not a soldier

#6 Rsm

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Posted 28 July 2010 - 06:22 AM

Bob1, you mention iPSCs. What experience, if any, do you have with these? I'm currently on a project looking at culturing Mouse Bone Marrow Cells and to expand the population of hematopoietic stem cells to then be able to treat them with IL-5 to generate an eosinophilic-like population of cells. I've had quite a lot of issues with getting the culture to work well, so maybe looking at/using iPSCs could be an option. What do you think?


I am not convinced that generating iPS from HSC and then differentiating them back to HSC makes much sense. Generating iPS is not "easy", you'll need at least 1 year to get it up and running. I think you should focus on HSC and discuss your problems with some specialists. iPS are extremely artificial, and you'll never know if success or failure of your culture system has anything to do with the exogeneous genes or chromatin landscape etc.
Sorry if I'm a bit negative.

Cheers,
Minna
I got soul, but I'm not a soldier

#7 bob1

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Posted 28 July 2010 - 12:59 PM

Bob1, you mention iPSCs. What experience, if any, do you have with these? I'm currently on a project looking at culturing Mouse Bone Marrow Cells and to expand the population of hematopoietic stem cells to then be able to treat them with IL-5 to generate an eosinophilic-like population of cells. I've had quite a lot of issues with getting the culture to work well, so maybe looking at/using iPSCs could be an option. What do you think?

Sorry, no experience with them at all, I've just seen the technology and thought it looked interesting.

I have never came across a feeder cell co-culture of cancer stem cells... Most people (at least in melanoma research) try to immediately inject them into mouse. And then it depends on the mouse, how "good" your CSC are. See for example http://www.ncbi.nlm....les/PMC2597380/ (Quintana 2008), hotly debated. I think most people agree that cell lines do not contain CSC. So you'd need access to fresh tumour samples, a FACS sorter and a severely immunocompromised mouse. I guess that'll be difficult.
Cheers,
Minna

It looks like you know much more than me, so I'll take your word for it. I was assuming that the processing through animals was just a test to show that they have tumour forming properties, and that they were also cultured separate from the animals.

#8 Rsm

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Posted 28 July 2010 - 11:51 PM

Sorry, I just realized that my link don't work. Click here.

Cheers,

Minna
I got soul, but I'm not a soldier

#9 ChaChi

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Posted 29 July 2010 - 05:25 AM

So, I guess I'm back to step 1...

Still doing research, if anyone has any ideas, please post.

I'm not allowed to use vertebrate animals, but I'm allowed to use invertebrates. That's just more detail in case anyone has any ideas.

Thanks,
ChaChi

#10 bob1

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Posted 29 July 2010 - 02:07 PM

You can do insect cell culture...

#11 ChaChi

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Posted 13 August 2010 - 12:02 PM

http://www.medicinen...ticlekey=112834

"Yeung and colleagues developed a new method of harvesting samples rich in cancer stem cells from bowel cancer cell lines and maintaining them in simple cell cultures in the lab. They used established cell lines, known as biological markers, to isolate the cancer stem cells, which were then collected and placed in largely standard cell culture conditions."

So from this article I understood that there are ways to do this without the fresh tumor or use of animals. Does anybody know anything more about this procedure?




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