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Human skeletal muscle cells culture?

Chen Linan chenlinan at 263.NET
Sat Mar 27 23:31:03 EST 1999


Dear Dr. Wolfgang,

In fact, in skeletal muscle from adult, there are some cells called satelite
cell who can divide something like precursor cell. I have beening culturing
some dishs of adult skeletal muscle cell. But several times, I failed to
culture others. The protocol is just the same. The only difference is the
skeletal muscle tissues come from different patients who undergo operations
of fracture. Any suggestion?

Best regards,

Chen Linan

Beijing Medical University


----------------------------------------------
Chen, 

> 1. How is it possible to culture human skeletal muscle cells from
> autopsy? Why can not I culture the human skeletul muscle cells from
> operation by enzyme dissociation methods?(Just repeat 3~5 times of
> 0.05% trysin-EDTA by 20 min each time). Oh, it is from a adult more
> than 40 years old.

Did you try collagenase instead? There should be a lot of connective 
tissue that trypsin hardly will be able to dissociate.

> 
> 2. When I culture the human skletal muscle cells of old people by
> tissue culture, sometimes it can grow out new cells, sometimes it
> can not, why?

skeletal muscle cells are said not to divide at all in adults. There 
should be suitable growth factors that could stimulate these cells to 
divide. (Try pubmed)

> 
> 3. AS about the enzyme dissociation methods, I want to know which is
> better for 0.05% trypsine-EDTA or 0.1% trpsine-collagenase?

probably the latter or collagenase alone

> 
> 4. How to design the experiment of the effect of bFGF on the
> proliferation of muscle cells without isotope method?

Incubate your cells with bromodeoxyuridine and stain the fixed cells 
with the antibody/peroxidase conjugate. Boehringer / Roche offers a 
pretty good kit for this purpose.

Some days ago, somebody posted here a method using a stain for the 
plasma membrane that is diluted by division. measured with a FACS 
analyzer. 



Cheers, 

Wo
> 
> 
> Best regards
> 
> Chen Linan
> 
> Beijing Medical University
> 
> 
> 
-----
usual disclaimers apply * This message is RNAse free - please don't touch!
-----                              
Wolfgang Schechinger   
Pathobiochemistry Dept.      
University of Tuebingen, Germany
email: wgschech at med.uni-tuebingen.de * wwWait:
http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
-------
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