Isolation of islets of Langerhans from the rat and pig pancreas using a modified UW solution from organ storage to islet purification.
The aim of this work was to determine the possibility of using the organ preservation solution UW for the enzymatic collagenase digestion step of the isolation of islets of Langerhans, although this solution is a calcium-free medium (collagenase being a calcium-dependent enzyme), high sodium and low potassium medium, which may be toxic for cells at the temperature needed for islet isolation. The rationale behind this work was that porcine islets are fragile, and that islets are poorly protected against oxidative stress: thus the cytoprotective properties of UW solution might be able to protect the islets during the digestion step, and improve the function of the isolated islets. As a first step, we verified that pancreas digestion with collagenase in a modified (i.e. hydroxyethyl starch deprived) UW solution was feasible: islets were randomly isolated from rat pancreas under either conventional conditions (usual ionic solution, IS), when the pancreas was inflated with UW solution (UW), and again when the pancreas was incubated for an additional hour (UW + 1) after inflation with UW and before the digestion step, thus simulating the time lapse between pancreas collection and islet isolation. In the last two conditions, collagenase was dissolved in UW solution for the digestion step.(ABSTRACT TRUNCATED AT 250 WORDS)
INSERM U 341, Service de Diabétologie, Hôtel-Dieu, Paris, France.
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Arbet-Engels
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Darquy
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D000241:Adenosine; D000493:Allopurinol; D000818:Animals; D002469:Cell Separation / Q000379:methods; D005978:Glutathione; D007328:Insulin; D007515:Islets of Langerhans / Q000166:cytology*; D008297:Male; D009926:Organ Preservation; D019188:Organ Preservation Solutions; D011887:Raffinose; D011897:Random Allocation; D051381:Rats / Q000033:anatomy & histology*; D013552:Swine / Q000033:anatomy & histology*; D014021:Tissue Preservation
D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D000493:Allopurinol; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose