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目的研究纳氏旋毛虫(Trichinellanelsoni,T7)肌幼虫细胞的分离和体外培养方法。方法消化法分离纳氏旋毛虫肌幼虫,用研磨法分离其细胞,倒置相差显微镜下观察分离细胞的形态并测量细胞核与细胞直径。用含10%胎牛血清的RPMI1640培养液进行细胞培养,并通过多重PCR对培养的细胞进行鉴定。结果纳氏旋毛虫肌幼虫细胞平均直径为11.72±1.13μm,细胞核平均直径为5.76±0.68μm。细胞在接种于培养液后24~72h开始贴壁生长,培养8~12d后单层细胞形成,贴壁细胞多为发亮、饱满、圆形细胞。对培养14d的细胞进行多重PCR分析,结果与纳氏旋毛虫肌幼虫具有相同的DNA条带。结论纳氏旋毛虫肌幼虫细胞在含10%胎牛血清的RPMI1640培养液中至少可存活20d。
Objective To study the isolation and cultivation of Trichinellanelsoni (T7) muscle larval cells. Methods The muscle larvae of Trichinella spiralis were isolated by digestion method and their cells were isolated by grinding method. The shapes of the cells were observed under inverted phase contrast microscope and the nuclei and cell diameters were measured. Cell culture was performed with RPMI1640 medium containing 10% fetal bovine serum, and the cultured cells were identified by multiplex PCR. Results The average diameter of Trichinella spiralis muscle larvae cells was 11.72 ± 1.13μm and the average diameter of nucleus was 5.76 ± 0.68μm. Cells were seeded in culture medium 24 ~ 72h after adherent growth began 8 to 12 days after the formation of monolayer cells, mostly adherent cells shiny, full, round cells. Multiplex PCR analysis of the cells cultured for 14 days showed the same DNA band as that of Trichinella spiralis muscle larvae. Conclusion Trichinella spiralis muscle larval cells can survive for at least 20 days in RPMI1640 medium containing 10% fetal bovine serum.