Li Na, Zhang Ying, Liu Yang, Xiu Zhilong, Tan Mingqian, Li Shen, Guo Xin, Wang Yu, Chen Li, Song Yizhe, Yu Weiting, Lv Guojun, Ma Xiaojun
3 University of Chinese Academy of Sciences, Beijing - China.
Int J Artif Organs. 2014 Jun;37(6):477-85. doi: 10.5301/ijao.5000335. Epub 2014 Jun 26.
Islet purification is usually performed using the density gradient separation method, but the purity of islets is low because exocrine cells and the embedded islets are hard to remove by using only the density gradient method. The aim of this study was to establish a new islet purification process comprising a hypertonic-hypotonic treatment step followed by a density gradient centrifugation step to improve the purity of islets.
The Plackett-Burman method was used to determine which factors had a significant influence on the purity of islets obtained after the hypertonic-hypotonic treatment step.
The hypertonic solution concentration and the incubation time were both found to have a significant effect on islet purity. The purity of islets obtained using the modified purification process was significantly higher than that of islets obtained by density gradient alone (97% vs. 87.23%). Importantly, good cell viability and normal insulin secretion ability of islets were maintained following the modified purification.
The new purification process allows isolation of islets with improved purity and does not compromise the viability or function of the islets.
胰岛纯化通常采用密度梯度分离法，但由于外分泌细胞和包埋的胰岛仅用密度梯度法难以去除，导致胰岛纯度较低。本研究的目的是建立一种新的胰岛纯化方法，该方法包括高渗-低渗处理步骤，随后进行密度梯度离心步骤，以提高胰岛纯度。
采用Plackett-Burman方法确定哪些因素对高渗-低渗处理步骤后获得的胰岛纯度有显著影响。
发现高渗溶液浓度和孵育时间均对胰岛纯度有显著影响。使用改良纯化方法获得的胰岛纯度显著高于仅通过密度梯度获得的胰岛纯度（97%对87.23%）。重要的是，改良纯化后胰岛保持了良好的细胞活力和正常的胰岛素分泌能力。
新的纯化方法能够分离出纯度更高的胰岛，且不影响胰岛的活力或功能。
