Dear Barry,

>  by providing us with some more information.

your right, but step by step otherwise we're stuck in one special topic,
which might bore the rest of the list, therefor I prefer to keep a wide
range of this topic (= different cell systems, as your already indicating
below).

> but many of the differentiation markers for cells become
>lost or at least inactive when cells are cultured.

This is where we try to manipulate, to keep/induce some adult properties.
I've  been dealing with this topic for 6 years and I can ensure that it's
still an exciting task.  

>1.    where are the muscle cells you are using from, species, strain,
location?
C2C12: hindlimb muscle of the mouse

>2.    are you separating cells or using pieces of tissue as in "organ
culture'?
It's on the plan to work with primary cultures, and then I'll have to
decide, but I think I'll go for cell monolayers.

>3.    if separating cells the technique used
enzymatic digestion.

>4.    the method of culture, is this stanadard, 3-dimensional culture etc.
>Barry
At the moment standard, but I'm open to alternatives.

Karim


>Karim Sultan wrote:
>
>> Hi All,
>> After recieving a countable number of answers for the cell culture request,
>> I'll try a different way:
>>
>> I'm interested to evaluate cell culture systems with morphological, incl.
>> immuno-methods, AND compare the results with the in vivo state. At the
>> moment I'm working with skeletal muscle cells. But the approach is similar
>> with other tissue.
>>
>> The problem is that pure "cell culture freaks" are fast in homogenizing
>> their cells and then the morphological  discussion finds a faster end.
>> Therefor I'm looking for Histo-Netters interested or working on topics
like:
>>
>> 1. adapting classical in vivo stains, like PAS, or
>> 2. dealing with culture conditions to gain more differentiation (e.g.
>> induce long Myotubes with marginal nuclei), or
>> 3. comparing differentiation markers.
>>
>> looking forward
>> Karim
>>
>> =============================================
>> Dr. Karim R. Sultan
>> Faculty of Veterinary Medicine VVDO
>> University of Utrecht
>> PO box 80.175
>> NL-3508 TD Utrecht
>>
>> Phone   (31) 30-2535361
>> Fax     (31) 30-2532365
>
>


=============================================
Dr. Karim R. Sultan
Faculty of Veterinary Medicine VVDO
University of Utrecht
PO box 80.175
NL-3508 TD Utrecht

Phone	(31) 30-2535361
Fax	(31) 30-2532365