Gotoh M, Ohzato H, Porter J, Maki T, Monaco A P
Second Department of Surgery, Osaka University Medical School, Japan.
Horm Metab Res Suppl. 1990;25:10-6.
We have been stressing the advantage of stationary digestion because of its simplicity and reproducible high yields of viable islets. In the present study optimal conditions of stationary in vitro collagenase digestion in mice and rats were examined. We also compared two possible routes for collagenase injection; ductal (PD) and portal venous (PV) based on subsequent islet yield and ability to reverse diabetes in rats. Three parameters which affect the quality of digestion are 1) collagenase concentration, 2) incubation time and 3) digestion temperature. Suitable conditions were easily determined and reproducible high yield of islets could be consistently obtained. The islets from one mouse pancreas (approximately 200 islets) could consistently restore normoglycemia in one STZ-induced diabetic mouse and the islets from one rat pancreas (500-600 islets) can restore normoglycemia in 5-6 STZ-induced diabetic mice within a couple of days. Islet yield in the PD method was greater than that in the PV method, and insulin release from PD islets in response to high glucose was well preserved after 24 hours of culture when compared to PV islets. The ability to restore normoglycemia in STZ-induced diabetic mice was well preserved when transplanting 100 PD islets as compared with the same number of PV islets. The PD islets revealed a well preserved structure with healthy endocrine cells, while the PV islets showed a dilated capillary network and distorted endocrine cell continuity. Histological examination of digested tissue following PD injection showed the complete destruction of pancreatic exocrine tissue, as well as mechanical separation and digestion of interstitial tissue between the islets and exocrine tissue, with the islet being preserved selectively intact.(ABSTRACT TRUNCATED AT 250 WORDS)
由于其操作简单且能稳定获得高产的活胰岛，我们一直强调静态消化的优势。在本研究中，我们考察了小鼠和大鼠静态体外胶原酶消化的最佳条件。我们还基于后续的胰岛产量和逆转大鼠糖尿病的能力，比较了两种胶原酶注射途径；即经导管（PD）和门静脉（PV）注射。影响消化质量的三个参数分别为：1）胶原酶浓度、2）孵育时间和3）消化温度。合适的条件易于确定，并且能够持续获得高产的胰岛。来自一只小鼠胰腺的胰岛（约200个胰岛）能够持续使一只链脲佐菌素诱导的糖尿病小鼠恢复正常血糖，而来自一只大鼠胰腺的胰岛（500 - 600个胰岛）能够在几天内使5 - 6只链脲佐菌素诱导的糖尿病小鼠恢复正常血糖。PD法的胰岛产量高于PV法，并且与PV胰岛相比，PD胰岛在培养24小时后对高糖刺激的胰岛素释放功能得到了良好保留。与相同数量的PV胰岛相比，移植100个PD胰岛时，其恢复链脲佐菌素诱导的糖尿病小鼠正常血糖的能力得到了良好保留。PD胰岛显示出结构良好，内分泌细胞健康，而PV胰岛则显示出扩张的毛细血管网络和内分泌细胞连续性受损。对PD注射后消化组织的组织学检查显示，胰腺外分泌组织被完全破坏，胰岛与外分泌组织之间的间质组织也被机械分离和消化，而胰岛则被选择性地完整保留。（摘要截选至250字）
