> 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
>>> Best regards
>> Chen Linan
>> Beijing Medical University
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