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目的观察旋毛虫幼虫在体外对正常小鼠肠上皮细胞(intestinal epithelial cells,IECs)的侵入及发育情况。方法将旋毛虫肌幼虫在小鼠胆汁中孵育2h,然后分别以不同密度(1.7条/cm2、4.2条/cm2及10.4条/cm2)接种不同类型培养基及不同面积培养板(9.6cm2和3.8cm2),在37℃5%CO2条件下培养,观察幼虫存活与发育情况。结果幼虫在半固体培养基+IECs中培养36h,发育至2～4期幼虫的百分率为30.93%,而在单纯半固体培养基与DMEM完全培养基+IECs中均不能发育为2～4期幼虫(P<0.01);半固体培养基+IECs中培养7d,低、中、高密度组虫体存活率分别为31.44%、7.82%和3.90%,差异有统计学意义(P<0.01)。低密度幼虫在两种培养板(9.6cm2和3.8cm2)培养7d,发育至成虫的百分率分别为33.29%和26.19%,差异无统计学意义(P>0.05),但仅在9.6cm2培养皿中观察到雄虫和受孕的雌虫。结论旋毛虫肌幼虫低密度接种大面积半固体培养基+IECs培养板培养,可发育至成虫。
Objective To observe the invasion and development of Trichinella spiralis larvae in normal mouse intestinal epithelial cells (IECs) in vitro. Methods Trichinella spiralis larvae were incubated in mouse bile for 2 hours, then inoculated with different culture medium and different culture plates (9.6cm2 and 3.8 respectively) with different density (1.7 / cm2, 4.2 / cm2 and 10.4 / cm2) cm2) and cultured in 5% CO2 at 37 ℃ to observe larval survival and development. Results The larvae grew in semi-solid medium + IECs for 36h, and the percentage of larvae which developed to 2-4 larvae was 30.93%. However, larvae did not develop 2-4 larvae in both semi-solid medium and DMEM complete medium + IECs (P <0.01). After cultured in semi-solid medium + IECs for 7 days, the survival rates of the larvae in low, medium and high density group were 31.44%, 7.82% and 3.90% respectively, with statistical significance (P <0.01). The low density larvae cultured in two culture plates (9.6cm2 and 3.8cm2) for 7 days and developed to adult percentage were 33.29% and 26.19%, respectively, with no significant difference (P> 0.05), but only in 9.6cm2 culture dish Male and female females were observed. Conclusion Trichinella spiralis larvae were inoculated into a large area of semi-solid culture medium + IECs culture plates to develop into adults.