当前培养方式对分离的胰岛质量的影响。

Influence of a current style of culture on the quality of isolated pancreatic islets.

作者信息

Takahashi H, Goto M, Ogawa N, Saitoh Y, Fujimori K, Kurokawa Y, Doi H, Satomi S

机构信息

Division of Advanced Surgical Science and Technology, Tohoku University, Sendai, Japan.

出版信息

Transplant Proc. 2008 Mar;40(2):358-9. doi: 10.1016/j.transproceed.2008.01.022.

DOI:10.1016/j.transproceed.2008.01.022

PMID:18374066

Abstract

OBJECTIVE

Comparable outcomes of islet transplantation with short periods of culture may be achieved with various culture media. To clarify the influence of a style of culture on isolated pancreatic islets, islet quality of fresh islets was compared with those cultured in several different fashions including not only for viability but also for inflammatory mediators.

MATERIALS AND METHODS

Wistar rat islets were cultured for 48 hours with CMRL including 10% allogeneic serum; CMRL including 0.5% human serum albumin (HSA); and Miami medium including 0.5% HSA. The influence of culture conditions on islet integrity was evaluated by survival rate of islets during culture and visual scoring. The influence of culture conditions on islet function and viability was examined by ADP/ATP tests, insulin/DNA content, and glucose stimulation tests.

RESULTS

Although the survival rates were similar for all groups, the visual scoring was lower in Miami medium. The stimulation index in glucose challenge tests was higher for fresh islets than the media (P = .02). Insulin/DNA ratios revealed the same tendency as glucose challenge tests (P = .0005). ADP/ATP ratio was lower in both the fresh and serum groups than in the others (P = .38), suggesting that apoptotic islets are relatively fewer in both fresh and serum groups. Most importantly, the expression of tissue factor (TF) on the islets was considerably lower in the fresh group, suggesting that a current style of culture could enhance TF-dependent instant blood-mediated inflammatory reactions after transplantation.

CONCLUSION

In conclusion, Isolated islets without prior culture shows characteristics beneficial for transplantation using current modes of culture.

摘要

目的

使用各种培养基或许能实现短期培养的胰岛移植的可比结果。为阐明培养方式对分离的胰岛的影响，将新鲜胰岛的质量与以几种不同方式培养的胰岛进行比较，不仅比较活力，还比较炎症介质。

材料与方法

将Wistar大鼠胰岛用含10%同种异体血清的CMRL培养基、含0.5%人血清白蛋白（HSA）的CMRL培养基以及含0.5% HSA的迈阿密培养基培养48小时。通过培养期间胰岛的存活率和视觉评分评估培养条件对胰岛完整性的影响。通过ADP/ATP测试、胰岛素/DNA含量和葡萄糖刺激试验检查培养条件对胰岛功能和活力的影响。

结果

尽管所有组的存活率相似，但迈阿密培养基中的视觉评分较低。葡萄糖激发试验中的刺激指数，新鲜胰岛高于培养基（P = 0.02）。胰岛素/DNA比值显示出与葡萄糖激发试验相同的趋势（P = 0.0005）。新鲜组和血清组的ADP/ATP比值均低于其他组（P = 0.38），表明新鲜组和血清组中凋亡胰岛相对较少。最重要的是，新鲜组胰岛上组织因子（TF）的表达显著较低，表明当前的培养方式可能会增强移植后TF依赖的即时血液介导的炎症反应。