LONG-TERM CULTURE OF HUMAN PANCREATIC-ISLETS IN AN EXTRACELLULAR-MATRIX - MORPHOLOGICAL AND METABOLIC EFFECTS
LONG-TERM CULTURE OF HUMAN PANCREATIC-ISLETS IN AN EXTRACELLULAR-MATRIX - MORPHOLOGICAL AND METABOLIC EFFECTS
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DOI：
10.1016/0303-7207(93)90046-m
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发表时间：
1993-07-01
影响因子：
4.1
通讯作者：
NICOL, M
中科院分区：
文献类型：
--
作者：
LUCASCLERC, C;MASSART, C;NICOL, M
本实验研究了人胰岛在I型胶原凝胶中包埋培养的各种条件，试图长期维持胰岛细胞的高度分化功能，特别是胰岛素分泌。通过胶原酶消化技术分离的胰岛细胞被涂在胶原凝胶上或胶原凝胶内，用Eagle最低必需培养基（5.5 mM d -葡萄糖）或RPMI 1640培养基（11 mM d -葡萄糖）补充10%的FCS和抗生素。两种培养基的比较表明，RPMI中培养的嵌入胰岛比MEM中培养的胰岛具有更高的基础胰岛素分泌率，存活时间更长，但与MEM中培养的胰岛相比，对急性血糖测试的反应受损。胰岛的分泌行为也与培养过程中发生的不同形态变化有关。直接嵌入胶原凝胶的胰岛或多或少保持其球形结构和最高的分泌能力。当被第二层胶原蛋白覆盖时，在胶原蛋白上生长的成熟的单层胰岛细胞逐渐完全重组为三维胰岛样结构，其分泌活性显著增强。两种培养物都能存活超过8周，从而证明了这种新模型对于长期培养的有用性。相比之下，在经过培养处理的塑料培养皿上的标准培养中，胰岛细胞迅速形成宽单层，表现出胰岛素分泌迅速而明确的下降，存活时间不超过14天。鉴于这些不同的培养条件，可能的机制负责激素释放紊乱和它们的意义在离体胰岛功能的体外研究进行了讨论。总之，这项工作是胶原基质对体外组织样结构建立或维持的允许作用的新例子，为长期培养的人类胰岛研究提供了新的模型。
In this experiment, various conditions for embedding cultures of human pancreatic islets in type I collagen gel were studied in an attempt to maintain the highly differentiated functions of islet cells and particularly insulin secretion over a long period of time. The islets isolated by a collagenase digestion technique were plated either on or within the collagen gel and refed with either Eagle's minimum essential medium (5.5 mM D-glucose) or RPMI 1640 medium (11 mM D-glucose) supplemented with 10% FCS and antibiotics. The comparison between the two culture media showed that embedded islets cultured in RPMI had a higher basal insulin secretion rate, survived longer than their MEM counterparts, but exhibited impaired response to an acute glucose test contrasting thus with islets cultured in MEM.The secretory behaviour of islets was also related to the different morphological modifications occurring during culture. Islets directly embedded within the collagen gel more or less maintained their spherical structure and highest secretory capacities. When overlaid with a second layer of collagen, well established monolayers of human islet cells grown on collagen underwent a gradual and complete reorganization into a three-dimensional islet-like structure with a striking reinforcement of their secretory activity. Both cultures were able to survive more than 8 weeks, thus proving the usefulness of such a new model for long-term culture. In contrast, standard cultures on culture treated plastic dishes on which islets cells rapidly established wide monolayers, exhibited a rapid and definitive decline in insulin secretion with a survival not exceeding 14 days.In the light of these different culture conditions, possible mechanisms responsible for disturbance of hormonal release and their implications for in-vitro study of isolated islets functions are discussed.In conclusion, this work is a new example of the permissive effects of collagen matrices on the establishment or maintenance of tissue-like structures in vitro, suggesting the definition of a new model for the study of human pancreatic islets in long-term culture.