Lerch M M, Hoppe-Seyler P, Matern S
Harvard Medical School, Beth Israel Hospital, Boston, MA 02215.
Biomed Biochim Acta. 1991;50(9):1115-25.
The use of pancreatic lobule preparations is one of the well-established approaches to study stimulus-secretion coupling in the exocrine pancreas in vitro. We have developed a kinetic system for the perfusion and intermittent incubation of rat pancreatic lobules. This model allows repeated hormone stimulation for up to 3 h while permitting rapid changes of the cellular environment with no accumulation of secretory or metabolic products. Tissue viability could be demonstrated over 6 h by in vivo toluidine blue exclusion, histology and electron microscopy. Lactate dehydrogenase leakage from cells over 6 h was only 2.4% of total content. No activated trypsin was detected in the perfusion medium. A biphasic dose response was established for cholecystokinin stimulation with a maximal response at 10(-8) M. We conclude that kinetic perfusion and incubation are technically feasible with rat pancreatic lobules. This in vitro model appears particularly suited for the investigation of pharmacologic and metabolic effects on the pancreatic acinar cell when rapid changes of the cellular environment are required and when the accumulation of secretory and metabolic products must be avoided. The technique described requires neither protease inhibition in the medium nor collagenase treatment of the cells.
使用胰腺小叶制剂是体外研究外分泌胰腺刺激-分泌偶联的成熟方法之一。我们开发了一种用于大鼠胰腺小叶灌注和间歇孵育的动力学系统。该模型允许重复激素刺激长达3小时，同时允许细胞环境快速变化，且无分泌或代谢产物积累。通过体内甲苯胺蓝排斥、组织学和电子显微镜可证明组织在6小时以上的活力。6小时内细胞乳酸脱氢酶泄漏仅占总含量的2.4%。在灌注培养基中未检测到活化的胰蛋白酶。建立了胆囊收缩素刺激的双相剂量反应，在10(-8) M时出现最大反应。我们得出结论，动力学灌注和孵育对大鼠胰腺小叶在技术上是可行的。当需要细胞环境快速变化且必须避免分泌和代谢产物积累时，这种体外模型似乎特别适合研究对胰腺腺泡细胞的药理和代谢作用。所述技术既不需要在培养基中抑制蛋白酶，也不需要对细胞进行胶原酶处理。
