| 邓磊玉林戈卢光琇.人胚胎干细胞支原体感染的检测和控制[J].现代生物医学进展英文版,2011,11(8):1451-1453. |
| 人胚胎干细胞支原体感染的检测和控制 |
| Examination and control of mycoplasma infection in humanembryonic stem cell culture |
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| DOI： |
| 中文关键词: 人胚胎干细胞 支原体 免疫荧光 抗支原体药物 |
| 英文关键词: human embryonic stem cell mycoplasma immunofluorence Plasmocin |
| 基金项目:国家高技术研究发展计划（863 项目）（2006AA02A102），国家重点基础研究发展计划（973 项目）
（2007CB948103），高等学校博士学科点专项科研基金（20090162110017） |
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| 中文摘要: |
| 目的：优化检测人胚胎干细胞支原体感染的方法,寻找控制支原体感染的途径。方法：利用hoeshest33258 染色检测感染支原
体的人胚胎干细胞，接触感染培养基的人胚胎成纤维细胞，比较两种方法检测效果；利用RNApolymeraseⅡ作为新的鉴定指标，
直接检测感染支原体的人胚胎干细胞；利用抗支原体药物对感染细胞进行处理，检测处理后细胞的感染状态。结果：
hoechest33258 染色后，受支原体感染人胚胎干细胞检测效果不明显，接触感染培养基的人胚胎成纤维细胞在培养7 天后有拉丝
状染色分布；RNApolymeraseⅡ染色则能直接检测出受感染的人胚胎干细胞表面粘附的支原体；利用抗支原体药物Plasmocin 对
感染细胞进行处理后hoechest33258 拉丝状染色基本消失，但持续培养后重新出现。结论：间接法使用hoechest33258 染色或者直
接利用RNApolymeraseⅡ染色都能够很好地检测人胚胎干细胞培养过程中的支原体感染。抗支原体药物Plasmocin 能够有效减
轻支原体感染情况，但是不能完全杀灭支原体。 |
| 英文摘要: |
| Objective: To optimize mycoplasma detection with immunofluorescence methods and control the mycoplasma
infection in human embryonic stem cell (hES) culture. Methods: Compare the two methods for mycoplasma detection by hoeshest33258,
one for direct staining of hES clones, another for indirect staining of feeder cells which touched hES culture medium sample. Took
RNApolymerase Ⅱ as a new detector for hES infection. Detect mycoplasma infection after Plasmocin treatment. Results: Hoechest33258
stanining showed linear stanining results when indirect sample were tested but there were no distinct results in direct staining sample.
RNApolymeraseⅡwas a good detector for mycoplasma infection. Linear stanining of hoeshest33258 almost disappeared after Plasmocin
treatment but appeared again in 2 months when withdrew the treatment. Conclusions: Indirect staining of hoeshest33258 and
RNApolymeraseⅡstaining are both available methods for mycoplasma detection. Plasmocin does not cure mycoplasma infection but
relieves it. |
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