The Effects of Cord Blood Serum on Survival of Rat Pancreatic Islets during In vitro Culturing

B. Karbek, Pınarlı Fa, G. Erden, E. Çakal, T. Delibaşı
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引用次数: 1

Abstract

Background: Transplantation of islets of Langerhans can improve glycometabolic control in type 1 diabetic patients; however, recipients usually require islets from 1-3 donor pancreas. Current isolation techniques generally recover <50% of the islets from a given pancreas. This study aimed to determine the effect of cord blood serum (CBS) on isolated pancreatic islet cells in vitro. Methods: Rat pancreatic islets were isolated via collagenase digestion, and then cultured with and without the presence of 50 μl, 100 μl, and 150 μl of CBS. Next, islet cell viability was determined using a fluorescein diacetate and propidium iodide mixture via fluorescence microscopy. Additionally, islet function was evaluated based on insulin secretion. Results: Treatment of primary cultures of rat pancreatic islets with CBS resulted in augmented β-cell viability. It was observed that the viability rate increased as the concentration of CBS increased and that viability was higher in the 150 μl of CBS group than in the control group. CBS was observed to preserve β-cell glucose responsiveness and insulin secretion under basal (3.3 mmoll-1) and stimulated (16.7 mmoll-1) glucose conditions. Moreover, a significant increase in insulin secretion was observed following treatment with 50 μl of CBS. Conclusions: The present findings show that cord blood serum treatment of freshly isolated rat islets had a positive effect on islet survival by increasing their viability. We think these findings suggest that CBS might be a suitable factor for optimizing and stimulating recovery, and subsequent function of islets during in vitro culturing, indicating that CBS might play an important role in the biophysiological function of pancreatic islets.
脐带血血清对体外培养大鼠胰岛存活的影响
背景:朗格汉斯胰岛移植可改善1型糖尿病患者的糖代谢控制;然而,受体通常需要来自1-3个供体胰腺的胰岛。目前的分离技术通常从给定的胰腺中回收不到50%的胰岛。本研究旨在探讨脐带血血清(CBS)对离体胰岛细胞的影响。方法:采用胶原酶消化法分离大鼠胰岛,分别添加50 μl、100 μl、150 μl CBS和不添加CBS培养大鼠胰岛。接下来,用荧光素和碘化丙啶混合物通过荧光显微镜测定胰岛细胞活力。此外,胰岛功能根据胰岛素分泌进行评估。结果:用CBS处理大鼠胰岛原代培养物可增强β细胞活力。结果表明,随着CBS浓度的增加,芽孢杆菌的存活率逐渐升高,且在CBS浓度为150 μl时,芽孢杆菌的存活率高于对照组。观察到CBS在基础(3.3 mmol -1)和刺激(16.7 mmol -1)葡萄糖条件下保持β细胞葡萄糖反应性和胰岛素分泌。此外,注射50 μl CBS后,胰岛素分泌明显增加。结论:脐带血血清处理对新分离的大鼠胰岛有积极的影响,可以提高胰岛的存活率。我们认为,这些结果表明,CBS可能是优化和刺激胰岛体外培养过程中恢复和后续功能的合适因素,表明CBS可能在胰岛的生物生理功能中发挥重要作用。
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