Successful nonfreezing, subzero preservation of rat liver with 2,3-butanediol and type I antifreeze protein.
Successful nonfreezing, subzero preservation of rat liver with 2,3-butanediol and type I antifreeze protein.
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DOI：
10.1006/jsre.2000.6053
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发表时间：
2001-03
期刊：
影响因子：
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通讯作者：
K. Soltys;A. Batta;B. Koneru
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文献类型：
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作者：
K. Soltys;A. Batta;B. Koneru
背景器官冷冻保存受到冰造成的损伤的阻碍。在零度以下的温度下不冷冻保存肝脏可能比目前的保存方法更有优势。方法SD大鼠随机分为3组。将UW肝脏（n = 6）储存在+4 ℃的威斯康星州大学（UW）溶液中。UWB肝脏（n = 6）在≤ 7 ℃下用UW +10%2，3-丁二醇离体灌注，并在-4 ℃下储存。AFP肝（n = 4）与UWB肝相同地保存，除了添加1 mg/ml的I型抗冻蛋白。24小时后，肝脏灌注与Krebs-Henseleit缓冲液（37 ℃）60分钟。胆汁生产，O（2）消耗（O（2）C），牛磺胆酸提取，和乳酸脱氢酶（LDH）的释放在灌注和肝脏腺嘌呤核苷酸含量和灌注结束时的能荷进行了测量。台盼蓝染色法检测细胞膜完整性。结果：在-4 ℃下保存的所有肝脏中，冰的形成都得到了防止。三组的胆汁生成、O（2）C和牛磺胆酸盐提取相似。储存在-4摄氏度的肝脏比储存在+4摄氏度的肝脏含有更多的腺嘌呤核苷酸，但能量电荷相似。AFP组的LDH释放量显著高于UWB和UW组（分别为63 vs 28和21 mU/min/g肝）（P < 0.05）。肝细胞和窦状隙细胞台盼蓝摄取在所有三组中相似。结论：在-4 ℃下保存的大鼠肝脏中，无论是否加用AFP，丁二酮都能有效防止冰形成长达24 h。尽管与当前的+4摄氏度方案一样有效，但在临床应用之前需要实现更长时间的零度以下保存。
BACKGROUND Organ cryopreservation is hindered by ice inflicted damage. Nonfreezing preservation of livers at subzero temperatures might offer advantages over current preservation. METHODS Sprague-Dawley rats were divided into three groups. UW livers (n = 6) were stored in University of Wisconsin (UW) solution at +4 degrees C. UWB livers (n = 6) were perfused ex vivo with UW + 10% 2,3-butanediol at < or =7 degrees C and stored at -4 degrees C. AFP livers (n = 4) were preserved identical to UWB livers, except for addition of 1 mg/ml of type I antifreeze protein. After 24 h livers were perfused with Krebs-Henseleit buffer (37 degrees C) for 60 min. Bile production, O(2) consumption (O(2)C), taurocholate extraction, and lactate dehydrogenase (LDH) release during perfusion and liver adenine nucleotide content and energy charge at the end of perfusion were measured. Cell membrane integrity was determined by trypan blue infusion. RESULTS Ice formation was prevented in all livers stored at -4 degrees C. Bile production, O(2)C, and taurocholate extraction were similar among three groups. Livers stored at -4 degrees C contained significantly more adenine nucleotides than livers stored at +4 degrees C but the energy charge was similar. LDH release was significantly greater (P < 0.05) in the AFP group vs UWB and UW (63 vs 28 and 21 mU/min/g liver, respectively). Hepatocyte and sinusoidal cell trypan blue uptake was similar in all three groups. CONCLUSIONS Butanediol with or without AFP was effective in preventing ice formation up to 24 h in rat livers stored at -4 degrees C. Although as effective as current +4 degrees C protocols, subzero preservation for longer periods needs to be achieved prior to clinical application.