大鼠胰岛分离纯化实验研究

A labotatory study of isolation and purification of islets of Langerhans from rat pancreas

目的介绍一种大鼠胰岛分离纯化方法。方法SD大鼠和Wistar大鼠共10只,以0.05%Ⅴ型胶原酶胆管灌注,经消化后用葡聚糖(dextran)不连续密度梯度法纯化胰岛。DTZ和AO/PI染色检测纯度及活性。结果胰岛收获量为(930±168)个,Ⅴ型胶原酶灌注及不连续密度梯度法可获得纯度高、活性好胰岛。结论以0.05%的胶原酶胆管灌注消化的方法可得纯度高、活性良好的胰岛。

Objective To introduce a procedure of isolation and purification of rat pancreatic islets of Langerhans. Methods 0.05 % collagenase type V 6-8ml was injected intraductally into ten SD and Wistar rats for digesting. After digestion, the preparation was purified by discontinuous dextran centrifugation. The viability and function in vitro of islets was determined by DTZ and AO/PI staining. Results The mean yields of islets of Langerhans per pancreas isolated by 0.05% collagenase were （930 ± 168） in per rat. Conclusion Collagenase type V in 0.05 % dextran digestion is a good method for isolation and purification of islets of Langerhans from rat pancreas.