Ultrastructural analyses of pancreatic grafts preserved by the two‐layer cold‐storage method and by simple cold storage in University of Wisconsin solution
Ultrastructural analyses of pancreatic grafts preserved by the two‐layer cold‐storage method and by simple cold storage in University of Wisconsin solution
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DOI：
10.1111/j.1432-2277.2002.tb00193.x
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发表时间：
2002-09
影响因子：
3.1
通讯作者：
Y. Iwanaga;Yusuyuki Suzuki;Y. Okada;H. Mori;I. Matsumoto;M. Mitsutsuji;Y. Tanioka;Y. Fujino;M. Tominaga;Y. Ku;Y. Kuroda
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文献类型：
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作者：
Y. Iwanaga;Yusuyuki Suzuki;Y. Okada;H. Mori;I. Matsumoto;M. Mitsutsuji;Y. Tanioka;Y. Fujino;M. Tominaga;Y. Ku;Y. Kuroda
摘要在犬胰腺保存模型中，采用威斯康星大学(UW)液的两层冷冻法(TLM)为移植胰腺提供了充足的氧气，使胰腺保存时间延长至96h。另一方面，UWM通过防止细胞肿胀将犬胰腺移植物保存长达72小时，这主要是因为它的高渗透压。本研究的目的是对这两种方法保存的犬胰腺移植物进行形态学分析，并探讨其不同的保存机制。犬胰腺经TLM保存72小时和96小时后(第1组和第3组)，UWM保存72小时和96h后(第2组和第4组)，用高效液相色谱法测定组织中的ATP含量，并用光镜和电子显微镜对移植物内成分进行详细的形态分析。用NIH图像1.62f软件计算单个线粒体和粗面内质网(PER)空泡化的平均面积。1组和3组组织中的ATP水平显著高于2组和4组(P<；0.05)。光镜下4组间差异无统计学意义。电子显微镜下，4组腺泡细胞均有不同程度的线粒体肿胀和粗面内质网空泡化。2组明显高于1组(P<；0.05)，4组明显高于3组(P<；0.05)。总之，TLM对细胞内细胞器、线粒体和粗面内质网具有良好的保护作用，保护时间可达72-96h。在TLM组中维持良好的移植物ATP水平可能会导致维持细胞内细胞器膜和细胞膜的完整性。
Abstract The two‐layer cold storage method (TLM) using University of Wisconsin (UW) solution supplies sufficient oxygen to pancreatic grafts during preservation and extends pancreas preservation time to up to 96h in the canine model. Simple cold storage in UW (UWM) on the other hand, preserves canine pancreas grafts for up to 72h by preventing cell swelling, mainly because of its high osmotic pressure. The aim of this study is to analyze morphologically dog pancreatic grafts preserved by these two methods with their different mechanisms. Immediately after preservation of canine pancreata by TLM for 72h and 96h (group 1 and group 3, respectively), and by UWM for 72h and 96h (group 2 and group 4, respectively), tissue ATP levels were determined using high‐performance liquid chromatography (HPLC), and detailed morphological analyses of intragraft components were performed using light‐ and electron microscopy. The mean areas of one mitochondrion and rough endoplasmic reticulum (PER) vacuolization were calculated by computer‐graphic analyses using NIH image 1.62f soft. The tissue ATP levels were significantly higher in groups 1 and 3 than groups 2 and 4 (P<0.05). Light microscopy demonstrated no marked difference among the 4 groups. By electron microscopy however, mitochondrial swelling and RER vacuolization were observed in acinar cells to various extents in the 4 groups. They were significantly more evident in group 2 than group 1 (P<0.05), and in group 4 than group 3 (P<0.05). In conclusion, TLM demonstrated excellent protection of intracellular organelles, mitochondria, and RER, up to 72–96h. Well‐maintained graft ATP levels in TLM groups may result in maintaining the integrity of intracellular organelle membranes as well as cellular membranes.