Noguchi Hirofumi, Naziruddin Bashoo, Shimoda Masayuki, Chujo Daisuke, Takita Morihito, Sugimoto Koji, Itoh Takeshi, Onaca Nicholas, Levy Marlon F, Matsumoto Shinichi
Baylor All Saints Medical Center, Baylor Research Institute, Fort Worth, TX, USA; †Institute of Biomedical Studies, Baylor University, Waco, TX, USA; ‡Department of Gastroenterological Surgery, Transplant and Surgical Oncology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.
†Institute of Biomedical Studies, Baylor University, Waco, TX, USA; §Baylor Regional Transplant Institute, Dallas and Fort Worth, TX, USA.
Cell Med. 2012 May 8;3(1-3):33-41. doi: 10.3727/215517912X639388. eCollection 2012 Jan.
For islet transplantation, islet purification minimizes the risks associated with islet infusion through the portal vein. However, islet purification may result in decreased numbers of islets recovered from digested tissue. In this study, we evaluated the effectiveness of performing supplemental purification (SP) after regular purification (RP). We designed the densities of low- and high-density solutions based on the outcome of RP. Moreover, a combined continuous osmolality/continuous density gradient for the SP was used in this study. Low-density/osmolality (1.075-1.110 g/cm/400-410 mOsm/kg) and high-density/osmolality (1.090-1.125 g/cm/495-505 mOsm/kg) solutions were produced by changing the volumetric ratio of iodixanol, 10 × HBSS, and RP solutions. The percentage of islet recovery (postpurification IE/prepurification IE × 100) after RP was 77.3 ± 5.6%, and the percentage of islet recovery after addition of SP was 85.3 ± 5.4%. In vitro and in vivo assessments showed that islet viability and function were not altered by the additional purification step. These data suggest that the addition of SP could contribute approximately 8% to islet recovery with viability and potency comparable to that obtained by RP and, therefore, that usage of the combined continuous density and continuous osmolality gradient for SP could efficiently improve islet equivalents in the final preparation.
对于胰岛移植，胰岛纯化可将经门静脉输注胰岛相关的风险降至最低。然而，胰岛纯化可能会导致从消化组织中回收的胰岛数量减少。在本研究中，我们评估了在常规纯化（RP）后进行补充纯化（SP）的有效性。我们根据RP的结果设计了低密度和高密度溶液的密度。此外，本研究中使用了用于SP的连续渗透压/连续密度梯度组合。通过改变碘克沙醇、10×HBSS和RP溶液的体积比来制备低密度/渗透压（1.075 - 1.110 g/cm³/400 - 410 mOsm/kg）和高密度/渗透压（1.090 - 1.125 g/cm³/495 - 505 mOsm/kg）溶液。RP后胰岛回收率（纯化后胰岛当量/纯化前胰岛当量×100）为77.3±5.6%，添加SP后的胰岛回收率为85.3±5.4%。体外和体内评估表明，额外的纯化步骤不会改变胰岛的活力和功能。这些数据表明，添加SP可使胰岛回收率提高约8%，其活力和效力与RP相当，因此，使用用于SP的连续密度和连续渗透压梯度组合可有效提高最终制剂中的胰岛当量。
