Hossein Aghdaie M, Azarpira N, Esfandiari E, Kaviani M, Golbabapour S, Shamsaeefar A, Kazemi K, Dehghani M, Bahador A, Salahi H, Nikeghbalian S, Malek-Hosseini S A, Geramizadeh B
Transplant Research Center, Shiraz University of Medical Sciences.
Department of Hepatobiliary Surgery, Shiraz University of Medical Sciences.
Int J Organ Transplant Med. 2019;10(3):108-113.
Liver transplantation is the only treatment for end-stage and genetic liver diseases. The main burden of this treatment is the shortage of both living and cadaveric liver donors. An alternative treatment is using liver cell transplantation, which can be obtained from unused livers for transplantation. These hepatocytes should be kept ready in viable and functional situation in a frozen state to be instantly used when they would be needed. In our previous experience, we had isolated hepatocytes from unused livers.
To find a preserving solution for increasing viability and function of the isolated hepatocytes that are stored to be transplanted.
9 cadaveric donor livers, which were not used for transplantation due to various causes such as severe steatosis, were selected to isolate hepatocytes. Various cold storage solutions were tried to find the best temperature for more viability and functionality for preservation of hepatocytes. University of Wisconsin (UW) solution and Williams E media were used as control media. 2 anti-apoptotic and anti-oxidative solutions, , α-lipoic acid and ursodeoxycholic acid (UDCA), were used as cold preservatives solutions. The numbers of viable hepatocytes were estimated by trypan blue method; the functionality was assessed by the cells ability to produce urea.
The highest number of viable and functional hepatocytes was obtained from freshly isolated cells. However, after preservation, the number of these viable hepatocytes and their functionality were not significantly different in cold storage solutions comparing to the control media used. Functionality of the isolated hepatocytes stored in UW with and without UCDA solution was similar to freshly isolated hepatocytes.
Preservatives with anti-apoptotic and antioxidant activity could not increase the number of viable hepatocytes. Functionality of cold storing hepatocytes could be preserved similar to freshly isolated hepatocytes by UW solution with and without UCDA.
肝移植是终末期和遗传性肝脏疾病的唯一治疗方法。这种治疗的主要负担是活体和尸体肝脏供体的短缺。一种替代治疗方法是使用肝细胞移植，肝细胞可从未使用过的用于移植的肝脏中获取。这些肝细胞应以冷冻状态保持存活和功能状态备用，以便在需要时立即使用。根据我们之前的经验，我们已从未使用过的肝脏中分离出肝细胞。
寻找一种保存溶液，以提高储存用于移植的分离肝细胞的活力和功能。
选择9个因各种原因（如严重脂肪变性）未用于移植的尸体供肝来分离肝细胞。尝试使用各种冷藏溶液，以找到保存肝细胞活力和功能的最佳温度。威斯康星大学（UW）溶液和威廉姆斯E培养基用作对照培养基。两种抗凋亡和抗氧化溶液，α-硫辛酸和熊去氧胆酸（UDCA），用作冷藏保存溶液。通过台盼蓝法估计活肝细胞的数量；通过细胞产生尿素的能力评估其功能。
从新鲜分离的细胞中获得的活肝细胞和功能肝细胞数量最多。然而，保存后，与对照培养基相比，冷藏溶液中这些活肝细胞的数量及其功能没有显著差异。储存在含和不含UCDA溶液的UW中的分离肝细胞的功能与新鲜分离的肝细胞相似。
具有抗凋亡和抗氧化活性的防腐剂不能增加活肝细胞的数量。含和不含UCDA的UW溶液可将冷藏肝细胞的功能保存得与新鲜分离的肝细胞相似。
