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

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。

Abstract:

[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.