DMEM培养基体外培养阴道毛滴虫效果的实验观察
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摘要
目的研究阴道毛滴虫在不同浓度胎牛血清DMEM培养基中的生长效果,比较阴道毛滴虫在DMEM培养基与15%肝浸汤培养基中的生长情况。方法调节DMEM培养基中胎牛血清终浓度分别为10%、20%、50%,接种密度为15×104/ml的阴道毛滴虫,置37℃恒温箱培养,每24小时取样一次,光镜观察虫体形态,利用血球计数板计数阴道毛滴虫增殖密度并计算增殖倍数。接种密度为1. 8×104/ml的阴道毛滴虫分别至DMEM、15%肝浸汤培养基中,比较阴道毛滴虫生长时间、增殖倍数及形态。结果接种密度为15×104/ml的阴道毛滴虫在终浓度为10%、20%、50%胎牛血清的DMEM培养基内分别培养48 h、24 h、72 h达增殖高峰。接种密度为1. 8×104/ml的阴道毛滴虫在DMEM培养基中120 h增殖倍数达高峰,为32. 63倍,虫体形态大而圆;在15%肝浸汤培养基中96 h增殖倍数达高峰,为59. 40倍,虫体形态多呈梨形;虫体在DMEM培养、肝浸汤培养基中生存时间最长分别为14 d、5 d。结论阴道毛滴虫在DMEM培养基中增殖率低于在肝浸汤培养基中,但虫体在DMEM培养基中存活时间更长,其形态大而圆,因此DMEM培养基适合阴道毛滴虫的体外生长,保种,染色,及形态观察。
Objective To investigate the proliferation of Trichomonas vaginalis in DMEM culture medium with different fetal bovine serum concentrations and compare the growth and proliferation of Trichomonas vaginalis between DMEM medium and 15%liver extract medium. Methods The 15 × 104/ml Trichomonas vaginalis were inoculated in the high glucose-DMEM medium with fetal bovine serum at final concentrations of 10%,20% and 50%,respectively. The proliferation of Trichomonas vaginalis was counted by blood cell count methods and the morphology of Trichomonas vaginalis was observed by light microscope every 24 hours. Trichomonas vaginalis was cultured in different media with initial inoculation of 1. 8 × 104/ml,the morphology,growth and proliferation of Trichomonas vaginalis in DMEM medium and 15% liver extracts were observed. Results Inoculation density of 15 × 104/ml Trichomonas vaginalis in DMEM medium with final serum concentrations of 10%,20%,50% reached the top of growth at 48 h,24 h,72 h. After inoculation of 1. 8 × 104/ml Trichomonas vaginalis in two different medium,Trichomonas vaginalis reached the top of growth at 120 hours,multiplied by 32. 63 folds in the DMEM medium,the morphology of organisms were uniform,large and round. it reached the top of growth at 96 hours,multiplied by 59. 40 folds in the 15% liver extract medium,the morphology of organisms were non-uniform,mostly pear-shaped. The survival time of the Trichomonas vaginalis in DMEM medium and 15% liver extracts can be up to 14 days,5 days,respectively. Conclusion Trichomonas vaginalis can grow well in the DMEM medium,in which they have longer survival time and their shape are more big,round than in the 15%liver extract medium,while its proliferation rate in the DMEM medium is lower than in the 15% liver extract medium. DMEM medium is the suitable medium for growth,teaching,staining and keeping species in laboratory.
Objective To investigate the proliferation of Trichomonas vaginalis in DMEM culture medium with different fetal bovine serum concentrations and compare the growth and proliferation of Trichomonas vaginalis between DMEM medium and 15%liver extract medium. Methods The 15 × 104/ml Trichomonas vaginalis were inoculated in the high glucose-DMEM medium with fetal bovine serum at final concentrations of 10%,20% and 50%,respectively. The proliferation of Trichomonas vaginalis was counted by blood cell count methods and the morphology of Trichomonas vaginalis was observed by light microscope every 24 hours. Trichomonas vaginalis was cultured in different media with initial inoculation of 1. 8 × 104/ml,the morphology,growth and proliferation of Trichomonas vaginalis in DMEM medium and 15% liver extracts were observed. Results Inoculation density of 15 × 104/ml Trichomonas vaginalis in DMEM medium with final serum concentrations of 10%,20%,50% reached the top of growth at 48 h,24 h,72 h. After inoculation of 1. 8 × 104/ml Trichomonas vaginalis in two different medium,Trichomonas vaginalis reached the top of growth at 120 hours,multiplied by 32. 63 folds in the DMEM medium,the morphology of organisms were uniform,large and round. it reached the top of growth at 96 hours,multiplied by 59. 40 folds in the 15% liver extract medium,the morphology of organisms were non-uniform,mostly pear-shaped. The survival time of the Trichomonas vaginalis in DMEM medium and 15% liver extracts can be up to 14 days,5 days,respectively. Conclusion Trichomonas vaginalis can grow well in the DMEM medium,in which they have longer survival time and their shape are more big,round than in the 15%liver extract medium,while its proliferation rate in the DMEM medium is lower than in the 15% liver extract medium. DMEM medium is the suitable medium for growth,teaching,staining and keeping species in laboratory.
引文
[1] DE BRUM VIEIRA P,TASCA T,SECOR WE. Challenges and persistent questions in treatment of trichomoniasis[J]. Curr Top Med Chem,2017,17(11):1249-1265.
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[3] OYETUNDE TS,CHELSEA NE. Frequency of Human Immunodeficiency Virus(HIV)in Trichomonas vaginalis Infected Women in Badagry,Lagos,Nigeria[J]. JRI,2016,17(1):61-63.
[4] KISSINGER P,AMEDEE A,CLARK RA,et al. Trichomonas vaginalis treatment reduces vaginal HIV-1 shedding[J]. Sexually Transm Dis,2009,36(1):11-16.
[5]王礼贤,龙溪,杨漪.阴道炎患者阴道毛滴虫感染状况调查[J].中国病原生物学杂志,2017,12(9):887-889.
[6]杨琚华,姚繁荣,答嵘,等.阴道毛滴虫在两种培养基中的生长与形态观察[J].中国寄生虫病防治杂志,2004,17(4):209-210.
[7]吴博文,苏盛通,陈惠亚,等.红枣牛肉汤与肝浸液培养阴道滴虫比较术[J].寄生虫与医学昆虫学报,2010,17(2):111-112.
[8]李兴武.不同浓度胎牛血清肝浸汤培养基培养阴道毛滴虫效果观察[J].中国病原生物学杂志,2009,4(10):800.
[9]赵建玲,高兴政.焦性没食子酸培养阴道毛滴虫方法的建立及其应用[J].中国血吸虫病防治杂志,2002,14(1):28-31.
[10]张潞渝,张静,叶彬,等.两种不同培养基体外培养阴道毛滴虫的增殖效果比较[J].重庆医学,2011,40(33):3389-3390.
[11]伏晨卉,张彦彦,苗玉倩,等.不同转种密度对体外培养阴道毛滴虫效果的影响[J].承德医学院学报,2015,32(6):536-537.
[2] CUENTHNER PC,SECOR WE,DEZZUTTI CS. Trichomonas vaginalis induced epithelial monolayer disruption and human immunodeficiency virus type 1(HIV-1)replication:implications for the sexual transmission of HIV-1[J]. Infect Immun,2005,73(7):4155-4160.
[3] OYETUNDE TS,CHELSEA NE. Frequency of Human Immunodeficiency Virus(HIV)in Trichomonas vaginalis Infected Women in Badagry,Lagos,Nigeria[J]. JRI,2016,17(1):61-63.
[4] KISSINGER P,AMEDEE A,CLARK RA,et al. Trichomonas vaginalis treatment reduces vaginal HIV-1 shedding[J]. Sexually Transm Dis,2009,36(1):11-16.
[5]王礼贤,龙溪,杨漪.阴道炎患者阴道毛滴虫感染状况调查[J].中国病原生物学杂志,2017,12(9):887-889.
[6]杨琚华,姚繁荣,答嵘,等.阴道毛滴虫在两种培养基中的生长与形态观察[J].中国寄生虫病防治杂志,2004,17(4):209-210.
[7]吴博文,苏盛通,陈惠亚,等.红枣牛肉汤与肝浸液培养阴道滴虫比较术[J].寄生虫与医学昆虫学报,2010,17(2):111-112.
[8]李兴武.不同浓度胎牛血清肝浸汤培养基培养阴道毛滴虫效果观察[J].中国病原生物学杂志,2009,4(10):800.
[9]赵建玲,高兴政.焦性没食子酸培养阴道毛滴虫方法的建立及其应用[J].中国血吸虫病防治杂志,2002,14(1):28-31.
[10]张潞渝,张静,叶彬,等.两种不同培养基体外培养阴道毛滴虫的增殖效果比较[J].重庆医学,2011,40(33):3389-3390.
[11]伏晨卉,张彦彦,苗玉倩,等.不同转种密度对体外培养阴道毛滴虫效果的影响[J].承德医学院学报,2015,32(6):536-537.