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joe7456

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Hi,

I am an undergraduate researcher at University of Georgia. I am currently assigned the task of taking mouse embryonic fibroblast (MEF) and coming up with a differentiation medium to culture the cells into various tissue. I would like to differentiate the MEF into cardiomyocytes or neuronal tissue. I have found two research articles which I will link below that have done this successfully, however, I am but a meek person and I need help understanding the methods and process of doing this.

The instructor I work with is wanting me to do this project independently. So I have to figure out what kind of media I need to use, all the components and growth factors that must be added. I must time the experiment right, use appropriate conditions, and see it through ( MEF ---> tissue).

Would anybody knowledgable in this area please give me some help or advice? I sure would appreciate it.

http://www.cell.com/action/showMethods?pii=S0092-8674(10)00771-3 (Cardiomyocytes)

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2829121/ (Neuronal cells)
 

SpartanWolverine

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You clearly are doing a good job finding background information on the project, but I think it's a bit much to assign an undergrad such a project as just a precursor to, I assume, some further research (as differentiation has been documented as possible for these strains). Your PI should be the knowledgeable one giving you advice.

I guess my point is... make sure your project has some sort of end game to it and that you're not just being used as someone to figure out how to make your PIs research happen. That should not be your role as an undergraduate RA unless it is what you signed on for.
 

joe7456

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I understand what you guys are saying. I am definitely stressed about the whole thing, but unfortunately my instructor is Asian and incredibly strict. I basically have to do all of this on my own. One of his research graduate assistants has said they have done MEF --> adipose tissue and MEF --> muscle tissue, however, they have not tried other differentiations. So I am basically kind of their puppet for now, and I understand it's a bit much, but I would still like to give it a try. I mean, someday people may benefit from this.
 

SurfingDoctor

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I understand what you guys are saying. I am definitely stressed about the whole thing, but unfortunately my instructor is Asian and incredibly strict. I basically have to do all of this on my own. One of his research graduate assistants has said they have done MEF --> adipose tissue and MEF --> muscle tissue, however, they have not tried other differentiations. So I am basically kind of their puppet for now, and I understand it's a bit much, but I would still like to give it a try. I mean, someday people may benefit from this.

Well that is a good attitude about it. Though I agree with SpartanWolverine, make sure you get something out of it too. Good luck.
 

moggat

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I have some experience with MEFs and it looks like you've got some good papers to start from. You should see if those labs have published more extensive write ups of their methods. For the most part, differentiation is basically tissue-> primary culture (MEFs)-> correct growth factors -> tissue you want, but it does vary from experiment to experiment. I think the bigger problem here is that your PI has given you no idea what the purpose of this is?? Differentiating tissue for flow cytometry vs. imaging vs. DNA analysis are all different, and until you know what it's going to be used for, it's going to be really hard to come up with a protocol, let alone execute it.

Are there any grad students in your lab who could help you go through the papers you've found or could shed some light on what your PI is planning to do? That should help you get a good starting point.

And as others have said ^^ this is kind of a crazy thing to expect from an undergrad; make sure you've understood whats really being expected of you and that you'll get something out of it.
 
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