Improvement of human islet cryopreservation by a p38 MAPK inhibitor
Improvement of human islet cryopreservation by a p38 MAPK inhibitor
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DOI：
10.1111/j.1600-6143.2007.01741.x
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发表时间：
2007-05-01
影响因子：
8.8
通讯作者：
Mullen, Y.
中科院分区：
文献类型：
--
作者：
Omori, K.;Valiente, L.;Mullen, Y.
p38丝裂原活化蛋白激酶（MAPK）的活化已显示引起用于移植的几个器官的缺血/再灌注损伤，并且在原发性胰岛移植物无功能中也起重要作用。p38 MAPK的激活也可能发生在胰岛冻存和解冻过程中。本研究将p38 MAPK抑制剂（p38 IH）应用于人胰岛冻存，以提高胰岛的冻存率和冻融后的质量。在无血清条件下，使用我们的标准程序冷冻保存、解冻和培养人胰岛。测试了三种类型的溶液：传统RPMI 1640培养基（RPMI）、新开发的胰岛冷冻保存溶液（ICS）和补充p38 IH、SD-282的ICS（ICS-p38 IH）。p38 MAPK的激活或抑制通过减少HSP 27底物的磷酸化来证明。ICS-p38 IH组解冻后第2天的胰岛恢复率最高，三组胰岛活力无显著差异。β细胞数量和功能在ICS-p38 IH冷冻保存的胰岛中最高。葡萄糖刺激的人C肽水平是86%的非冷冻胰岛移植到NODscid小鼠后4周测量。这一改进可能为建立胰岛库提供机会，并允许将冷冻保存的胰岛用于临床移植。
The activation of p38 mitogen-activated protein kinase (MAPK) has been shown to cause ischemia/reperfusion injury of several organs used for transplantation and also to play a significant role in primary islet graft nonfunction. Activation of p38 MAPK may also occur during islet cryopreservation and thawing. In this study, a p38 MAPK inhibitor (p38IH) was applied to human islet cryopreservation to improve islet yield and quality after thawing. Under serum-free conditions, human islets were cryopreserved, thawed and cultured using our standard procedures. Three types of solutions were tested: conventional RPMI1640 medium (RPMI), a newly developed islet cryopreservation solution (ICS), and ICS supplemented with a p38IH, SD-282 (ICS-p38IH). Activation or inhibition of p38 MAPK was demonstrated by the diminished phosphorylation of HSP27 substrate. Islet recovery on day 2 after thawing was highest with ICS-p38IH and islet viability was not significantly different in the three groups. beta Cell numbers and function were the highest in islets cryopreserved with ICS-p38IH. Glucose-stimulated human C-peptide levels were 86% of that of the nonfrozen islets when measured 4 weeks after transplantation into NODscid mice. This improvement may provide an opportunity to establish islet banks and allow the use of cryopreserved islets for clinical transplantation.