Explant culture - All method of cell isolation coming from animal tissue (Jan/11/2007 )

Hi,

for a future study, I will need to make explant culture of aorta... Indeed, I must analyze the vascular smooth muscle cells. But I don't know the principle of this method. Other and last question for this post, what is the difference between the explant culture and the use of collagenase and elastase????????

Thanks for your reply!!!!!

Good research

Boumbo_doc

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To me "explant culture" is when you culture pieces of aorta and let cells grow from it, and "collagenase, elastase" is when you digest the pieces of aorta before culturing it.

For mice vSMCs, collagenase method seems to work better than explant culture. One of my colleague had very nice cultures with this method.

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[/quote]
To me "explant culture" is when you culture pieces of aorta and let cells grow from it, and "collagenase, elastase" is when you digest the pieces of aorta before culturing it.

For mice vSMCs, collagenase method seems to work better than explant culture. One of my colleague had very nice cultures with this method.
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Thank you very much,
but which are the avantanges and disadvantages of these two methods?

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Things to think about :

a) It is always easier to go for large animal Aorta's which will give you more tissue to work with and therefore give you more cells for experiments.

a) Once cells are in culture their phenotype changes from Contractile to Synthetic

Explant SMC take longer to grow (upto 7-10 days), but you can achieve a "purer" culture.

c) Enzyme digest allows SMC for experiments immediately, but are prone to Fibroblast contamination.

d) Which ever method you choose, YOU MUST ALWAYS CHARACTERISE YOUR CELLS i.e. SM alpha actin staining.


Commercial companies now supply VSMC which have been isolated and characterised, such as Promocell, Biowhittaker and Clonetics, but are relatively expensive.

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