一种制备新生隐球菌单细胞的方法  

Using cell sorting techniques to explore the recovery of individual Cryptococcus neoformans cells

作　　者：周利艳 付钰[1,2] Liyan Zhou;Yu Fu(State Key Laboratory of Microbial Resources,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101,China;Savaid Medical School,University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]中国科学院微生物研究所微生物资源前期开发国家重点实验室,北京100101 [2]中国科学院大学存济医学院,北京100049

出　　处：《微生物学报》2019年第2期326-333,共8页Acta Microbiologica Sinica

基　　金：国家自然科学基金(31571288);中国科学院跨学科创新团队;英国皇家学会的牛顿先进奖学金(NA140085)~~

摘　　要：【目的】建立流式细胞仪分选新生隐球菌单细胞的方法,确定新生隐球菌单细胞恢复生长的条件和能力。【方法】利用Moflo XDP流式细胞分析分选仪,体外测定不同条件下新生隐球菌恢复生长的比率。【结果】建立了流式细胞仪新生隐球菌单细胞分选流程。确定流式细胞仪分选得到的新生隐球菌单细胞具有恢复生长的能力,恢复生长的能力受营养条件和菌株差异的影响。在营养丰富的条件下,新生隐球菌JEC21和H99单细胞恢复生长比率分别为74%和89%。在寡营养条件下,JEC21和H99单细胞恢复生长比率分别为37%和80%。JEC21生长比率随细胞数的增加而升高,细胞数为100个时,生长比率为55%;细胞数为1000个时,生长比率为97%。【结论】流式细胞仪分选得到新生隐球菌单细胞具有恢复生长的能力,生长能力受营养条件、菌株差异的影响。[Objective]A method of sorting single cells of Cryptococcus neoformans by flow cytometry was established to determine factors influencing proliferation of single C.neoformans cells.[Methods]The Moflo XDP flow cytometric sorter was used.The clonal recovery rate of C.neoformans cell was determined under various conditions.[Results]Single C.neoformans cells sorting process by cytometry was established.Single C.neoformans cells could recover growth which was affected by nutritional conditions and strains.Under nutrient-rich conditions,the clonal recovery rates of C.neoformans stains JEC21 and H99 were 74%and 88%,respectively.Under oligotrophic conditions,the clonal recovery rate of JEC21 was 37%;while the clonal recovery rate of H99 was 80%.The clonal recovery rate of JEC21 increased with increasing initial number of cells.When the cell number of JEC21 strain was 100,the clonal recovery rate was 55%;when the cell number was 1000,the clonal recovery rate was 97%.[Conclusion]The single C.neofroman cells sorted by flow cytometry had the capacity of recovery to growth,and the capacity was affected by nutritional conditions and strains.

关 键 词：单细胞分选技术 流式细胞仪 新生隐球菌 

分 类 号：R379.5[医药卫生—病原生物学]