制冷技术

本文以新鲜的乳腺癌组织为研究对象，采用程序降温盒搭配-80℃冰箱的降温方式，检测样本相对活性、免疫组化等指标，与直接放入液氮和直接放入-80℃冰箱2种降温方式相对比，分析了程序降温盒搭配-80℃冰箱的降温方式对组织的影响，使用复温后的组织进行肿瘤浸润淋巴细胞（TIL）的组织培养。结果表明：-80℃冰箱中程序降温法的保存效果最好，组织相对活性在55%以上，同时搭配5种低温保护剂的第7组效果最佳，细胞存活率为64%，液氮组的细胞存活率最低为32%;3种低温保存方法均未使组织形态发生明显变化。-80℃冰箱冻存的2种方法在复温后均可培养出TIL细胞，冻存后的组织培养出的细胞数量均小于新鲜组的40%，表型为CD3+、CD8+的细胞比例较少，但细胞活率较高，均在80%以上。

Fresh breast cancer tissues are used as the research subjects in this paper. A programmed cooling box combined with an-80 °C freezer is employed for sample freezing, while relative activity, immunohistochemical staining, and other indicators are measured. This method is compared with two alternative approaches: direct immersion in liquid nitrogen and direct placement in an-80 °C freezer. The impact of the programmable cooling box in conjunction with the-80 °C freezer on tissue is analyzed, and the rewarmed tissues are used for tumorinfiltrating lymphocyte(TIL) tissue culture. The results show that the preservation effect of the programmable cooling method at-80 °C freezer is the best, with tissue relative activity above 55%. Among them, the seventh group, which is simultaneously paired with five cold preservation agents, show the best effect at cell survival rate of 64%, while the relative activity of the liquid nitrogen group is the lowest at 32%. None of the three lowtemperature preservation methods cause significant changes in tissue morphology. Both methods of storage in the-80 °C freezer can culture TIL cells after rewarming, with the cell quantity cultured from the stored tissues being less than 40% of the fresh group. The phenotype of the cultured cells is predominantly CD3+ and CD8+, with a relatively low proportion, but the cell viability is high, all above 80%.