Miyagi-Shiohira Chika, Kobayashi Naoya, Saitoh Issei, Watanabe Masami, Noguchi Yasufumi, Matsushita Masayuki, Noguchi Hirofumi
Department of Regenerative Medicine, Graduate School of Medicine, University of the Ryukyus , Okinawa , Japan.
† Okayama Saidaiji Hospital , Okayama , Japan.
Cell Med. 2016 Sep 1;9(1-2):45-51. doi: 10.3727/215517916X693131. eCollection 2017 Jan 8.
Islet purification is one of the most important steps of islet isolation for pancreatic islet transplantation. The most common method of islet purification is density gradient centrifugation using a COBE 2991 cell processor. However, this method can damage islets mechanically through its high shearing force. We recently reported that a new purification method using large plastic bottles effectively achieves a high yield of islets from the porcine pancreas. In the present study, we evaluated the methods of making a continuous density gradient. The gradient was produced with a gradient maker and two types of candy cane-shaped stainless steel pipes. One method was to use a "bent-tipped" stainless steel pipe and to load from a high-density solution to a low-density solution, uploading the stainless steel pipe. The other method was to use a regular stainless steel pipe and to load from a low-density solution to a high-density solution, leaving the stainless steel pipe in place. There were no significant differences between the two solutions in terms of the islet yield, rate of viability or purity, score, or the stimulation index after purification. Furthermore, there were no differences in the attainability or suitability of posttransplantation normoglycemia. Our study shows the equivalency of these two methods of islet purification.
胰岛纯化是胰岛移植中胰岛分离最重要的步骤之一。最常见的胰岛纯化方法是使用COBE 2991细胞处理器进行密度梯度离心。然而，这种方法会因其高剪切力而机械损伤胰岛。我们最近报道了一种使用大塑料瓶的新纯化方法，能有效地从猪胰腺中获得高产率的胰岛。在本研究中，我们评估了制备连续密度梯度的方法。该梯度是用梯度制备仪和两种拐杖糖形状的不锈钢管产生的。一种方法是使用“弯尖”不锈钢管，从高密度溶液向低密度溶液加载，将不锈钢管向上移。另一种方法是使用普通不锈钢管，从低密度溶液向高密度溶液加载，将不锈钢管留在原位。在胰岛产量、活力率或纯度、评分或纯化后的刺激指数方面，两种溶液之间没有显著差异。此外，移植后正常血糖的可实现性或适宜性也没有差异。我们的研究表明这两种胰岛纯化方法是等效的。
