Baylor All Saints Medical Center, Baylor Research Institute, Fort Worth, TX, USA.
Cell Transplant. 2012;21(2-3):493-500. doi: 10.3727/096368911X605402.
For pancreatic islet transplantation, the most common method of islet purification is density gradient centrifugation because of the differences in density between islets and acinar tissue. The density of islets/acinar tissue depends on several conditions, such as osmolality of purification solution. In this study, we evaluated the osmolality of iodixanol-controlled density gradients (400, 450, and 500 mOsm/kg) on the islet purification step. The density of the purification solutions was controlled by changing the volumetric ratio of iodixanol and the purification solutions (iodixanol-Kyoto solutions; IK solutions). The osmolality of density gradients was controlled by addition of 10× Hanks balanced salt solution (HBSS) solution. Density of both islets and acinar tissue increased relative to increase of the osmolality of purification solutions. There were no significant differences among the three groups on islet yield after density-adjusted purification and the rate of postpurification recovery. In vitro and in vivo assays suggest that the quality of islets was similar among the three groups. Our data suggest that efficacy of purification and quality of isolated islets is similar when the osmolality of purification solutions is between 400 and 500 mOsm/kg and density adjustment is applied. Since the density of islet and acinar tissue is changed according to osmolality, the density adjustment is important when using several osmolality solutions.
对于胰岛移植，胰岛纯化最常用的方法是密度梯度离心，因为胰岛和腺泡组织的密度存在差异。胰岛/腺泡组织的密度取决于几个条件，如纯化溶液的渗透压。在这项研究中，我们评估了碘克沙醇控制的密度梯度（400、450 和 500 mOsm/kg）在胰岛纯化步骤中的渗透压。通过改变碘克沙醇和纯化溶液（碘克沙醇-京都溶液；IK 溶液）的体积比来控制纯化溶液的密度。通过添加 10×Hanks 平衡盐溶液（HBSS）溶液来控制密度梯度的渗透压。胰岛和腺泡组织的密度都随纯化溶液渗透压的增加而增加。在经过密度调整的纯化后，三组之间的胰岛产量和纯化后回收率没有显著差异。体外和体内试验表明，三组之间胰岛的质量相似。我们的数据表明，当纯化溶液的渗透压在 400 和 500 mOsm/kg 之间且进行密度调整时，纯化的效果和分离的胰岛的质量是相似的。由于胰岛和腺泡组织的密度根据渗透压而变化，因此在使用几种渗透压溶液时，密度调整很重要。
