Human skeletal muscle cells culture?

Wolfgang Schechinger Wolfgang.Schechinger at med.uni-tuebingen.de
Sat Mar 27 12:04:19 EST 1999


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
> 
> 
> 
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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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