玻璃化冷冻技术在新鲜胃癌样本深低温保存中的应用  

Application of vitrification in cryopreservation for fresh gastric cancer samples

作　　者：亓垚 吴珊珊 李姣姣 谷凯 赵子君 宋嫔 段逸凡 蔚想霞 乔天麟 郑文彦 陈天宝 张小燕[1,2] 谭元生 QI Yao;WU Shanshan;LI Jiaojiao;GU Kai;ZHAO Zijun;SONG Pin;DUAN Yifan;WEI Xiangxia;QIAO Tianlin;ZHENG Wenyan;CHEN Tianbao;ZHANG Xiaoyan;TAN Yuansheng(National Engineering Center for Biochip at Shanghai,Shanghai 201203,China;Shanghai Outdo Biotech Co.,Ltd,Shanghai 201203,China;Jihua Low Temperature Biotechnology(Guangdong)Co.Ltd,Foshan 528000,China)

机构地区：[1]生物芯片上海国家工程研究中心,上海201203 [2]上海芯超生物科技有限公司,201203 [3]季华低温生物科技(广东)有限公司,佛山528000

出　　处：《中国医药生物技术》2025年第S1期1-9,共9页Chinese Medicinal Biotechnology

基　　金：上海市专业技术服务平台建设(19DZ2295000);佛山高新区高技术产业化创业扶持项目(2320197000077);上海工程技术研究中心建设项目(19DZ2283100)。

摘　　要：目的探讨人胃癌和癌旁组织经过玻璃化冷冻及复苏后对类器官培养及相关组织形态学和标志物表达特征的影响。方法将4例人胃癌和癌旁组织各自分为新鲜样本组和玻璃化冷冻复苏组,各组样本在类器官接种前进行细胞计数和活率检测,类器官培养过程中也对各组相同代数及培养时间的类器官进行计数和活率检测,类器官培养完成后进行HE染色及Ki67、CEA、MUC6和CK19等标志物的IHC检测。结果经过玻璃化冷冻复苏后的组织消化后细胞计数和活率有一定程度的下降;接种后形成的类器官数量较少,生长速度略慢,但一部分样本在传代之后,逐渐恢复与新鲜组织培养的类器官类似的生长速度;并且在组织形态学和标志物表达水平方面与新鲜组织培养的类器官基本一致。结论玻璃化冷冻体系经过优化和调整后,可以成为长期保存组织样本活性的有效手段。Objective To investigate the effects of vitrification cryopreservation and recovery on organoid culture,as well as the histomorphological and marker expression characteristics in human gastric cancer and adjacent tissues.Methods Gastric cancer and adjacent tissue samples from 4 patients were divided into fresh sample groups and vitrification cryopreservation-recovery groups.Before organoid seeding,cell counting and viability testing were performed for each group.During organoid culture,organoids of the same passage and culture time from each group were also counted and tested for viability.Histomorphologic feature and expression of markers,such as Ki67,CEA,MUC6,and CK19,were observed by HE staining and immunohistochemical(IHC)detection.Results Compared to fresh tissues,the cell count and viability of tissues after vitrification cryopreservation and recovery showed a certain degree of decline.The number of organoids formed after seeding was lower,and their growth rate was also slightly slower.However,in some samples,the growth rate gradually recovered to a level similar to that of organoids cultured from fresh tissues after passaging.Moreover,the histomorphology and marker expression levels of these organoids were largely consistent with those of organoids cultured from fresh tissues.Conclusion After optimization and adjustment,the vitrification cryopreservation system can serve as an effective method for long-term preservation of tissue sample viability.

关 键 词：类器官 生物样本活库 胃癌及癌旁组织 玻璃化冷冻 

分 类 号：Q-34[生物学]