Cold preservation of islets in UW solution - With special reference to apoptosis
Cold preservation of islets in UW solution - With special reference to apoptosis
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DOI：
10.1016/j.jss.2005.10.006
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发表时间：
2006-06-15
影响因子：
2.2
通讯作者：
Inoue, Kazutomo
中科院分区：
文献类型：
--
作者：
Shirouzu, Yasumasa;Gu, Yuanjun;Inoue, Kazutomo
背景。胰岛细胞凋亡的进展。关于低温保存下的细胞凋亡，我们所知甚少。我们研究了胰岛在威斯康星大学溶液中的生存能力和功能。材料和方法。取离体大鼠胰岛进行隔夜培养（隔夜组），然后在RPMI 1640培养基中37℃培养7 d（培养组）或在UW溶液中4℃保存7 d（保存组）。采用葡萄糖刺激胰岛素分泌试验评价。采用tdt介导的dutp -生物素缺口末端标记法（TUNEL）检测细胞凋亡。采用逆转录聚合酶链反应（RT-PCR）检测caspase mRNA的表达及Bax与Bcl-2的比值。培养组7 d胰岛恢复率显著低于保存组（44.0 +/- 3.7% vs 75.0 +/- 4.9%, P < 0.05）。培养组刺激指数显著低于过夜组（2.1 +/- 0.2 vs 4.1 +/- 0.4, P < 0.05）。培养组细胞凋亡指数显著高于隔夜组和保存组（38.0 +/- 3.0% vs 10.8 +/- 2.0和27.0 +/- 4.0%,P < 0.05）。Caspase 3、8、9 mRNA在培养组的表达高于其他各组。培养组细胞Bax/Bcl-2水平明显低于过夜组（3.2 +/- 0.66 vs 8.1 +/- 0.95, P < 0.05），提示细胞分离后早已发生凋亡。保存组的恢复和功能优于培养组。细胞凋亡导致了胰岛细胞在培养条件下的损失，而在低温保存条件下则明显受到抑制。(c) 2006爱思唯尔公司版权所有。
Background. Apoptosis progresses in cultured islets. Little is known with regard to apoptosis under cold preservation. We examined viability and function of islets in University of Wisconsin (UW) solution.Materials and methods. Isolated rat islets were cultured overnight (overnight group) and further treated with 7-day culture in RPMI 1640 medium at 37 degrees C (culture group) or 7-day preservation in UW solution at 4 degrees C (preservation group). They were evaluated by glucose-stimulated insulin secretion test. Apoptosis was examined by TdT-mediated dUTP-biotin nick end-labeling (TUNEL) assay. Expression of caspase mRNA and the ratio of Bax to Bcl-2 were evaluated by reverse-transcriptase polymerase chain reaction (RT-PCR).Results. Islet recovery after 7 days was significantly lower in culture group than in preservation group (44.0 +/- 3.7% versus 75.0 +/- 4.9%, P < 0.05). The stimulation index in the culture group was significantly lower than in the overnight group (2.1 +/- 0.2 versus 4.1 +/- 0.4, P < 0.05). The apoptotic index in the culture group was significantly higher than both in the overnight group and in the preservation group (38.0 +/- 3.0% versus 10.8 +/- 2.0 and 27.0 +/- 4.0%, P < 0.05). Caspase 3, 8, and 9 mRNA in the culture group expressed more than in the other groups. Bax/Bcl-2 in the culture group was significantly lower than in the overnight group (3.2 +/- 0.66 versus 8.1 +/- 0.95, P < 0.05), suggesting that apoptosis had been already destined early after isolation.Conclusions. The preservation group showed better recovery and function than the culture group. Apoptosis contributed to islet loss under culture and it was significantly suppressed under cold preservation. (c) 2006 Elsevier Inc. All rights reserved.