PREPARATION OF, AND STUDIES ON, FREE CELL-SUSPENSIONS FROM MOUSE PANCREATIC-ISLETS
PREPARATION OF, AND STUDIES ON, FREE CELL-SUSPENSIONS FROM MOUSE PANCREATIC-ISLETS
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DOI：
10.1007/bf01221634
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发表时间：
1974-01-01
期刊：
影响因子：
8.2
通讯作者：
LERNMARK, A
中科院分区：
文献类型：
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作者：
LERNMARK, A
从肥胖高血糖小鼠胰腺中分离胰岛，制备游离胰岛细胞悬液。在含有1 mM EGTA的无Ca2+ hepes缓冲Krebs-Ring - er培养基中，通过机械摇动10秒破坏100个批次的胰岛。从200-500个胰岛中，大约可以在悬浮中获得2.2×106cells，根据细胞和胰岛中的DNA含量计算，产量约为55%。分离的胰岛细胞在光镜和电子显微镜下保存完好，并且通过活细胞计数、放射性溶解测定和胰岛素释放来判断，似乎表现出高度的活力。胰岛素从胰岛细胞悬浮液中释放，按每个细胞、每个DNA或胰岛素含量或每个填充细胞体积计算，葡萄糖单独或与茶碱联合刺激。与完整的离体胰岛相比，葡萄糖反应较低。
Pancreatic islets, isolated from the pancreas of obese-hyperglycemic mice, were used to prepare free islet cells in suspension. Batches of 100 islets were disrupted by mechanical shaking for 10 sec in a Ca2+-free HEPES-buffered Krebs-Ring'er medium containing 1 mM EGTA. From 200–500 islets, about 2.2×106cells could be obtained in suspension, corresponding to a yield of roughly 55% as calculated from the content of DNA in cells and islets. The isolated islet cells appeared well-preserved in the light and the electron microscope and seemed to exhibit a high degree of viability as judged by viable cell counts, a radioactive assay for lysis and insulin release. Insulin release from islet cell suspensions, as calculated per cell, per content of DNA or insulin or per packed cell volume, was stimulated by glucose alone or in combination with theophylline. The glucose response was low compared with that of intact isolated islets.