The Experimental Study On Experession Of Nestin And Ngn3 Positive Cells In Different Islet Purity In Rats

Objective:To investigate the Nestin and Ngn3 positive cells in the donor islets of different purity in rats.Methods:30 male SD rats(islet donors) were randomly divided into 3 groups. Islets of pancreas were isolated by pancreatic ductal injection of collagenase, then discontinuous gravity gradient purification (Ficoll-400 ) was used to obtain three different islets purity . GroupI(N=10): Dissociated cell preparations were not purified,Group II(N=10): Islets sediments were purified with Ficoll (25%) gradient density,Group III(N=10): Islet sediments were purified with Ficoll (25%,11%) gradient density. The levels of protein of Nestin and Ngn3 positive cells were detected by immunohistochemistry.The expression of Nestin and Ngn3 mRNA was amplified by reverse transcription- Polymerase chain reaction(RT-PCR).Results:1. Three different islets purity were produced by continuous digestion and different purified methods, groupI: 43.6±6.29%,group II: 65.3±4.40%,group III: 77.6±6.36%. There were significant difference among various group(P﹤0.05). Besides,three kinds of islet yields were...