Reciprocal stimulation of pancreatic acinar and stellate cells in a novel long-term in vitro co-culture model
Reciprocal stimulation of pancreatic acinar and stellate cells in a novel long-term in vitro co-culture model
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DOI：
10.1016/j.pan.2016.03.012
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发表时间：
2016-07-01
期刊：
影响因子：
3.6
通讯作者：
Laukkarinen, Johanna
中科院分区：
文献类型：
--
作者：
Blauer, Merja;Laaninen, Matias;Laukkarinen, Johanna
背景/目的：胰腺星状细胞（Pancreatic stellate cells, PSCs）是胰腺中主要的纤维化细胞。已知腺泡细胞损伤可触发PSC激活。为了便于实验分析腺泡细胞与PSCs之间的串扰，我们建立了一种长期共培养的体外系统。材料和方法：从小鼠胰腺中获得在体外长期培养中能保持分泌表型的PSCs和腺泡细胞。建立24孔双室共培养模型，孔中接种腺泡细胞，组织培养插入PSCs。腺泡细胞- 3t3成纤维细胞共培养作为对照。维持4天后，分析腺泡室细胞形态、分泌能力、坏死（HMGB1）、凋亡（TUNEL）和炎症（NF κ B）。分析PSCs的迁移活性和细胞外基质（ECM）蛋白表达。结果与平行单一培养进行了比较。结果：单培养和与成纤维细胞共培养的腺泡细胞表现出健康的单层排列，并且能够通过增加淀粉酶释放来响应0.1 nM的蛋白刺激。与PSCs共培养使腺泡细胞形态发生显著变化，使其对促分泌剂刺激不敏感。在共培养的腺泡细胞中发现NF κ B活化和坏死改变，但未发现凋亡。共培养提高了PSCs的迁移活性和ECM蛋白的表达。结论：共培养的腺泡细胞和PSCs之间的体液相互作用相互影响其细胞功能。其两个可分离的细胞区室，共培养系统提供了一个多功能的培养设置，允许独立操作和分析两种细胞类型。（三）2016年IAP和EPC。Elsevier B.V.版权所有。
Background/objectives: Pancreatic stellate cells (PSCs) are the key fibrogenic cells in the pancreas. Acinar cell injury is known to trigger PSC activation. To facilitate the experimental analysis of the crosstalk between acinar cells and PSCs, an in vitro system for their long-term co-cultivation was developed.Materials and methods: PSCs and acinar cells capable of retaining their secretory phenotype in long-term in vitro culture were obtained from mouse pancreata. A dual-chamber co-culture model was built in 24 well format with acinar cells seeded in the wells and PSCs in tissue culture inserts. Acinar cell-3T3 fibroblast co-cultures served as controls. After 4-day maintenance, the acinar compartment was analyzed for cell morphology, secretory capability, necrosis (HMGB1), apoptosis (TUNEL) and inflammation (NF kappa B). PSCs were analyzed for migratory activity and extracellular matrix (ECM) protein expression. The results were compared to parallel monocultures.Results: Acinar cells in monoculture and in co-culture with fibroblasts exhibited a healthy monolayer arrangement and an ability to respond to 0.1 nM caerulein stimulus by increased amylase release. Co-culture with PSCs caused marked changes in acinar cell morphology and rendered them insensitive to secretagogue stimulus. Activation of NF kappa B and necrotic changes, but not apoptosis, were identified in co-cultured acinar cells. Co-culture increased the migratory activity and ECM protein expression of PSCs.Conclusions: Humoral interactions between acinar and PSCs in co-culture were shown to reciprocally affect their cellular functions. With its two separable cell compartments the co-culture system provides a versatile culture setting that allows independent manipulation and analysis of both cell types. (C) 2016 IAP and EPC. Published by Elsevier B.V. All rights reserved.