国际医学寄生虫病杂志
國際醫學寄生蟲病雜誌
국제의학기생충병잡지
INTERNATIONAL JOURNAL OF MEDICAL PARASITIC DISEASES
2014年
4期
227-229,233
,共4页
弓形虫%速殖子%人巨噬细胞%人包皮成纤维细胞%体外培养
弓形蟲%速殖子%人巨噬細胞%人包皮成纖維細胞%體外培養
궁형충%속식자%인거서세포%인포피성섬유세포%체외배양
Toxoplasma gondii%Tachyzoites%Human macrophage%Human foreskin fibroblasts%In vitro culture
目的 比较弓形虫RH株速殖子在人巨噬细胞和人包皮成纤维细胞内的增殖情况. 方法 按照弓形虫速殖子∶细胞为1∶1的比例将弓形虫RH株速殖子分别与人巨噬细胞和人包皮成纤维细胞共培养0、6、12、24、48 h后,于显微镜下观察细胞内虫体增殖情况.在两种细胞培养瓶中分别接种弓形虫RH速殖子,连续传代,每隔10代取弓形虫速殖子,将其密度调整为1×107个/ml,0.3 ml/只,腹腔接种ICR健康小鼠,记录小鼠存活时间. 结果 在相同条件下弓形虫速殖子侵入人巨噬细胞的时间早于侵入人包皮成纤维细胞的时间;共培养48 h时,绝大部分弓形虫速殖子胀破细胞,游离在细胞外.在两种细胞培养瓶中分别接种3×106弓形虫RH速殖子,约72 h后均可获得大量游离的弓形虫速殖子,2～3d传1代.人巨噬细胞内连续传代约30次,每代可获得(1×107～4×107)个速殖子,约增殖3～13倍;人包皮成纤维细胞中连续传代30次,每代可获得(1×107～3×107)个速殖子,约增殖3～10倍.弓形虫速殖子传代次数为10、20和30代时,人巨噬细胞内传代的弓形虫速殖子接种小鼠的存活时间分别为(4.3±0.5)、(4.0±0.8)、(4.3±1.2)d,人包皮成纤维细胞内传代的弓形虫速殖子接种小鼠的存活时间分别为(4.0±0.0)、(3.7±0.9)、(4.3±0.5)d,各代之间差异无统计学意义(P＞0.05),弓形虫速殖子毒力未见明显改变. 结论 弓形虫RH株速殖子可在人巨噬细胞和人包皮成纤维细胞内增殖并长期稳定传代.
目的 比較弓形蟲RH株速殖子在人巨噬細胞和人包皮成纖維細胞內的增殖情況. 方法 按照弓形蟲速殖子∶細胞為1∶1的比例將弓形蟲RH株速殖子分彆與人巨噬細胞和人包皮成纖維細胞共培養0、6、12、24、48 h後,于顯微鏡下觀察細胞內蟲體增殖情況.在兩種細胞培養瓶中分彆接種弓形蟲RH速殖子,連續傳代,每隔10代取弓形蟲速殖子,將其密度調整為1×107箇/ml,0.3 ml/隻,腹腔接種ICR健康小鼠,記錄小鼠存活時間. 結果 在相同條件下弓形蟲速殖子侵入人巨噬細胞的時間早于侵入人包皮成纖維細胞的時間;共培養48 h時,絕大部分弓形蟲速殖子脹破細胞,遊離在細胞外.在兩種細胞培養瓶中分彆接種3×106弓形蟲RH速殖子,約72 h後均可穫得大量遊離的弓形蟲速殖子,2～3d傳1代.人巨噬細胞內連續傳代約30次,每代可穫得(1×107～4×107)箇速殖子,約增殖3～13倍;人包皮成纖維細胞中連續傳代30次,每代可穫得(1×107～3×107)箇速殖子,約增殖3～10倍.弓形蟲速殖子傳代次數為10、20和30代時,人巨噬細胞內傳代的弓形蟲速殖子接種小鼠的存活時間分彆為(4.3±0.5)、(4.0±0.8)、(4.3±1.2)d,人包皮成纖維細胞內傳代的弓形蟲速殖子接種小鼠的存活時間分彆為(4.0±0.0)、(3.7±0.9)、(4.3±0.5)d,各代之間差異無統計學意義(P＞0.05),弓形蟲速殖子毒力未見明顯改變. 結論 弓形蟲RH株速殖子可在人巨噬細胞和人包皮成纖維細胞內增殖併長期穩定傳代.
목적 비교궁형충RH주속식자재인거서세포화인포피성섬유세포내적증식정황. 방법 안조궁형충속식자∶세포위1∶1적비례장궁형충RH주속식자분별여인거서세포화인포피성섬유세포공배양0、6、12、24、48 h후,우현미경하관찰세포내충체증식정황.재량충세포배양병중분별접충궁형충RH속식자,련속전대,매격10대취궁형충속식자,장기밀도조정위1×107개/ml,0.3 ml/지,복강접충ICR건강소서,기록소서존활시간. 결과 재상동조건하궁형충속식자침입인거서세포적시간조우침입인포피성섬유세포적시간;공배양48 h시,절대부분궁형충속식자창파세포,유리재세포외.재량충세포배양병중분별접충3×106궁형충RH속식자,약72 h후균가획득대량유리적궁형충속식자,2～3d전1대.인거서세포내련속전대약30차,매대가획득(1×107～4×107)개속식자,약증식3～13배;인포피성섬유세포중련속전대30차,매대가획득(1×107～3×107)개속식자,약증식3～10배.궁형충속식자전대차수위10、20화30대시,인거서세포내전대적궁형충속식자접충소서적존활시간분별위(4.3±0.5)、(4.0±0.8)、(4.3±1.2)d,인포피성섬유세포내전대적궁형충속식자접충소서적존활시간분별위(4.0±0.0)、(3.7±0.9)、(4.3±0.5)d,각대지간차이무통계학의의(P＞0.05),궁형충속식자독력미견명현개변. 결론 궁형충RH주속식자가재인거서세포화인포피성섬유세포내증식병장기은정전대.
Objective To compare the proliferation of Toxoplasma gondii RH strain tachyzoites in human macrophage and human foreskin fibroblasts.Methods Toxoplasma gondii RH strain tachyzoites were inoculated into human macrophage and human foreskin fibroblasts according to the rate of 1 ∶ 1,the proliferation of tachyzoites was observed under microscope at 0,6,12,24,48 hour after inoculation.At the same time,the long-term in vitro culture of Toxoplasma gondii tachyzoites maintained in human macrophage and human foreskin fibroblasts was established.The Toxoplasma gondii RH strain tachyzoites were taken out at every 10 generations then inoculated intraperitoneally into ICR mice according to 3 ×106 tachyzoites per mice.Then the survival days of the mice were observed.Results The time of Toxoplasma gondii tachyzoites invading into human macrophage is earlier than that into human foreskin fibroblasts.At 48 hour after co-culture,most of the two cells were destroyed by tachyzoites.In the long-term culture system,a large amount of free tachyzoites could be obtained after 72 h incubation.In human macrophage,(1×107～4×107 tachyzoites could be obtained each generation for 30 generations in continually culture with the proliferation rate of 3-13 times while 1 ×107～3×107 tachyzoites obtained each generation for 30 generation in HFF cells with the proliferation rate of 3-10 times.So the proliferation of tachyzoites was stable in both cells.The survival time of the mice inoculated with tachyzoites at 10,20 and 30 generation cultured in human macrophages was (4.3+ 0.5) d,(4.0+0.8) d and (4.3+1.2) d respectively,while it was (4.0+0.0) d,(3.7+0.9) d and (4.3+0.5) d respectively for the mice infected with tachyzoites cultured in HFF cells.The difference among various generations was not significant statistically (P＞0.05) and their virulence to mice showed no decrease(P＞0.05).Conclusion The proliferation of Toxoplasma gondii RH strain tachyzoites cultured in human macrophage and human foreskin fibroblasts was stable for long-term generations.
