Basic Study on Preservation Conditions Of Donor's Lungs In Lung Transplantation - Development of The Standard Method for Evaluating Alveolar Type II Cells With Their Surfactant Synthetic Activities As An Indicator -

需要进行肺移植，以使许多肺细胞功能得到很好的保存。到目前为止，这一时间限制在6小时左右。不幸的是，由于这一时间限制，大多数患者并未获救。如果开发出新的保存液或更好的保存条件，肺移植将为更多的患者开放。在这项研究中，我们打算利用分离的大鼠肺泡II型细胞建立评估新的保存液或更好的保存条件的标准方法。采用Mason等人描述的标准方法从大鼠肺中分离II型细胞。分离的II型细胞在标准培养液(D-MEM)和两种保存液(胞外型Perfodex(P)液和胞内型Euro-Collins(EC)液)中与表面活性物质脂质前体-~3H胆碱孵育20h。肺表面活性物质合成活性(~3H-胆碱对~3H-磷脂酰胆碱的摄取)在P液中的II型细胞高于D-MEM和EC液。分离后，用相同的方法测定表面活性物质的合成活性。P液保存的大鼠肺组织中的II型细胞数明显多于EC液保存的细胞。而肺泡表面活性物质的合成活性在两种类型的肺泡灌洗液中是相似的，因此，胞外型P液作为肺泡表面活性物质的功能标记物更适合于肺保存。

Lung transplantation needs to be performed so that many of lung cell functions are well preserved. This time limit has been around 6 hours so far. Unfortunately, most of the patients has not been saved because of this time limit. If new preservation fluids or better preservation conditions are developed, lung transplantation will be opened for more patients. In this study, we intended to develop the standard methods for evaluating new preservation fluids or better preservation conditions using rat isolated alveolar type II cells.Type II cells were isolated from rat lungs using a standard method described by Mason, et el. Isolated type II cells were incubated for up to 20 hrs with ^3H choline, a precursor of surfactant lipid both in the standard medium (D-MEM) and in two 'kinds of preservation fluids; extracellular type Perfodex (P) solution and intracellular type Euro-Collins (EC) solution. Surfactant synthetic activity (uptake of ^3H-choline to ^3H-phoshatidylcholine) was greater in type II cells in P solution than in those in D-MEM and EC solutions.Next, we isolated type II cells from rat lungs which were perfused with P and EC solutions and preserved in the same solutions for 6 hours at 4 degree. After isolation, surfactant synthetic activities were estimated with the same methods. The number of type II cells isolated from rat lungs preserved in P solution was greater than that in EC solution. However, surfactant synthetic activity was similar in both type II cells.In conclusion, the extracellular type P solution was suitable in lung preservation in a view of surfactant synthetic activity as a functional marker of type II cells.