摘要
[目的]比较单独及混合胶原酶消化小鼠乳腺癌移植瘤组织后获取的细胞活性。[方法]用4T1细胞建立小鼠乳腺癌动物模型,待成瘤后取乳腺癌肿瘤组织。比较4种消化酶(Ⅰ型胶原酶、Ⅳ型胶原酶、Ⅰ+Ⅳ型混合胶原酶和商用胶原酶),两种消化方法:单酶或多酶单次消化30 min;单酶或多酶分次消化30 min。用碘化丙啶(PI)染色各组消化后的细胞悬液,流式细胞仪检测细胞活率。[结果]不同酶单次消化后的细胞活率分别为:Ⅰ型胶原酶为55. 5±7. 2%、Ⅳ型胶原酶为26. 9±7. 6%、Ⅰ+Ⅳ型混合胶原酶为43. 5±10%、商用胶原酶为30. 4±10. 6%;在分次消化后分别为:Ⅰ型胶原酶为70. 6±4. 1%、Ⅳ型胶原酶为65. 5±17. 2%,Ⅰ+Ⅳ型混合胶原酶为78. 3±2. 2%,商用胶原酶为48. 5±11. 6%。[结论]使用Ⅰ+Ⅳ型混合胶原酶分次消化得到的乳腺癌移植瘤细胞活性最高,并为小鼠乳腺癌移植瘤组织消化提供了一种有效的新方法。
[Objective]To compare the cell viability of mouse breast cancer transplanted tumor tissue digested with alone and mixed collagenase.[Method]The mouse breast cancer animal model was established with 4 T1 cells.The cell viabilities after digested by four digestive enzymes(collagenase typeⅠ,collagenase typeⅣ,mixed collagenase typeⅠandⅣand commercial collagenase),compared in two digestion methods:single or multiple enzyme to single digestion for 30 min;single or multiple enzyme to multiple digestion for 30 min.The cell viability by flow cytometry was stained with propidium iodide(PI).[Result]In the single or multiple enzyme to single digestion group,the cell viability after digested by collagenase type I was 55.5±7.2%,the ones by collagenase typeⅣwas 26.9±7.6%,the ones by mixed collagenase typeⅠandⅣwas 43.5±10%,the ones by commercial collagenase was 30.4±10.6%,respectively.In the single or multiple enzyme to multiple digestion group,the cell viability after digested by collagenase typeⅠwas 70.6±4.1%,the ones by collagenase typeⅣwas 65.5±17.2%,the ones by mixed collagenase typeⅠandⅣwas 78.3±2.2%,and the ones by commercial collagenase was 48.5±11.6%,respectively.[Conclusion]The highest cell viability of breast cancer transplanted tumor cells was harvested by the mixed collagenase typeⅠandⅣmultiple digestion,and it provided an effective new method for the digestion of breast cancer xenograft tissue in mouse.
作者
郭婷婷
王方芳
黄巧容
陈正举
孟文彤
GUO Ting-ting;WANG Fang-fang;HUANG Qiao-rong;CHEN Zheng-ju;MENG Wen-tong(West China Hospital,Sichuan University,Chengdu 610041,China)
出处
《生物技术》
CAS
2019年第4期387-390,共4页
Biotechnology
关键词
乳腺癌
胶原酶
消化
细胞活性
breast cancer
collagenase
digest
cell viability
