I am trying to isolate a specific cell in the eye but I don't know how to digest the matrix components without lysing or destroying the cells. The matrix is primarily made of collagen, GAGs and proteoglycans.
Hi
It is highly dependent on what kind of tissue you are going to digest because ECM from different tissues is varying in type of collagen and based on that different collagenase (such as type I,II,II or A and D) must be into account. I, personally, digest adipose tissue to isolated mesenchymal stem cells by collagenase type I because this enzyme mainly digest collagen type that is present in soft tissue such as adipose tissue or kidney. Collagenase type III would be nice for ESs isolation from endometrium and for the last example type D is most appropriate for cartilage digestion in companion with dispase I or II.
Best
Hi Swetha
You can look at digestion protocols for cartilage as a starting point as cartilage has collagen and gag. Generally such tissues are digested using collagenase.
Please go over a chapter number 20 of this book I am enclosing, hope that helps. If any question, e-mail directly to Dr. Matt Hilton directly, he is in Duke Univ. Nice person.
Here you are
Article Manual Isolation of Adipose-derived Stem Cells from Human Li...
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