What are the best known methods (processes) for immortalizing human somatic cells?
Need references and vendors for necessary equipment and reagents. |
Clarification of Question by
xn1-ga
on
11 Apr 2003 15:18 PDT
pls provide details of procedure(s) if they are not explained in the references.
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Request for Question Clarification by
pafalafa-ga
on
11 Apr 2003 17:45 PDT
I can certainly direct you to the major references on immortalization
techniques, and the articles themselves (many of which are accessible
online) have highly detailed "Methods" sections that explain the
processes used, and identify the key techniques, reagents, and
equipment.
For instance, a typical "Methods" paragraph from a major paper on
immortalization looks like this:
"The cells were then incubated for 1 h with a 1:200 dilution of
fluorescein isothiocyanate-conjugated donkey anti-mouse IgG secondary
antibody (Jackson Laboratories, Bar Harbor, ME). All antibody
dilutions
were made in PBS. The slides were mounted in FA Mounting Buffer (Bio-
Rad, Hercules, CA) and coverslipped. Photographs were taken with a
Zeiss
microscope (Carl Zeiss MicroImaging Inc., Thornwood, NY) equipped for
epifluorescence."
Would this information suffice as an answer?
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Clarification of Question by
xn1-ga
on
12 Apr 2003 09:45 PDT
(1) techniques needs to be applicable to human somatic cells.
(2) ideally, there should be an initial comparison to other techniques
and why the method(s) you outline is best (i.e. because it's simpler,
cheaper, more reliably repeatable).
(3) ideally, there should be an explanation of why each step is taken,
what is being accomplished and why is it done this way. (i.e. what's
the point of incubating it for 1 h? and at what temperature(s)?).
Describe incubator if it's unusual. Taking the photo sounds like
effort to document - but is not a needed part of the process?
(4) ideally a lay person should be able to understand it or follow up
the references to get a better idea of what the stuff is and where it
comes from and even how that's done (i.e. PBS?)
(5) If that's all there is to it, maybe there should be an
explanation of what's needed to perpetuate the culture and/or to store
it. I mean, how do I know these guys are immortal now.
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Clarification of Question by
xn1-ga
on
01 May 2003 12:29 PDT
Already understood. I'd be intrigued by research with the various
immortalization methods applied to stem cells, reintroduction into the
individual (host?) and what the results were.
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Clarification of Question by
xn1-ga
on
01 May 2003 12:41 PDT
The various techniques, when applied to human cells, which chromosomes
are affected and how?
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