脱氧胆酸对C57BL/6小鼠回肠类器官生长的影响
目的：建立小鼠小肠类器官培养系统，探讨脱氧胆酸（DCA）对小肠类器官生长的影响。方法取8周龄C57BL/6小鼠末段回肠，EDTA法分离，收集隐窝后Matrigel基质胶包埋，于完整培养基中培养。以无水酒精和正常小肠类器官为对照，予100μmol/L高浓度DCA短期干预2 d，10μmol/L DCA长期干预10 d，并去除DCA后再长期培养10 d，观察类器官的生长和出芽情况。结果高浓度DCA短期干预类器官，肠道类器官球形结构形成率、肠道类器官结构[形成率、类器官演化率和出芽数量均明显减少（P〈0.05），短期去除DCA四项指标仍减少（P〈0.05），长期去除DCA后仅肠道类器官结构形成率和出...
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| Published in | 南方医科大学学报 Vol. 37; no. 1; pp. 6 - 12 |
|---|---|
| Main Author | |
| Format | Journal Article |
| Language | Chinese |
| Published |
南京医科大学附属上海一院临床医学院消化科，上海 200080
2017
上海交通大学附属第一人民医院 国际医疗保健中心，上海 200080 上海市胰腺疾病重点实验室，上海 200080%上海交通大学附属第一人民医院 消化科，上海 200080 上海市胰腺疾病重点实验室，上海 200080 上海交通大学附属第一人民医院 消化科，上海 200080 上海市胰腺疾病重点实验室，上海 200080%南京医科大学附属上海一院临床医学院消化科，上海 200080 |
| Subjects | |
| Online Access | Get full text |
| ISSN | 1673-4254 |
| DOI | 10.3969/j.issn.1673-4254.2017.01.02 |
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| Abstract | 目的：建立小鼠小肠类器官培养系统，探讨脱氧胆酸（DCA）对小肠类器官生长的影响。方法取8周龄C57BL/6小鼠末段回肠，EDTA法分离，收集隐窝后Matrigel基质胶包埋，于完整培养基中培养。以无水酒精和正常小肠类器官为对照，予100μmol/L高浓度DCA短期干预2 d，10μmol/L DCA长期干预10 d，并去除DCA后再长期培养10 d，观察类器官的生长和出芽情况。结果高浓度DCA短期干预类器官，肠道类器官球形结构形成率、肠道类器官结构[形成率、类器官演化率和出芽数量均明显减少（P〈0.05），短期去除DCA四项指标仍减少（P〈0.05），长期去除DCA后仅肠道类器官结构形成率和出芽数量减少（P〈0.05）。低浓度DCA短期干预肠道类器官结构形成率和出芽数量减小（P〈0.05），长期干预肠道类器官球形结构形成率、肠道类器官结构形成率和出芽数量明显减少（P〈0.05），去除DCA后仅肠道类器官结构形成率和出芽数量低于正常（P〈0.05）。结论本研究成功建立小鼠小肠类器官培养技术，DCA损伤小肠类器官并影响其生长，长期去除DCA后可部分恢复其功能。 |
|---|---|
| AbstractList | 目的：建立小鼠小肠类器官培养系统，探讨脱氧胆酸（DCA）对小肠类器官生长的影响。方法取8周龄C57BL/6小鼠末段回肠，EDTA法分离，收集隐窝后Matrigel基质胶包埋，于完整培养基中培养。以无水酒精和正常小肠类器官为对照，予100μmol/L高浓度DCA短期干预2 d，10μmol/L DCA长期干预10 d，并去除DCA后再长期培养10 d，观察类器官的生长和出芽情况。结果高浓度DCA短期干预类器官，肠道类器官球形结构形成率、肠道类器官结构[形成率、类器官演化率和出芽数量均明显减少（P<0.05），短期去除DCA四项指标仍减少（P<0.05），长期去除DCA后仅肠道类器官结构形成率和出芽数量减少（P<0.05）。低浓度DCA短期干预肠道类器官结构形成率和出芽数量减小（P<0.05），长期干预肠道类器官球形结构形成率、肠道类器官结构形成率和出芽数量明显减少（P<0.05），去除DCA后仅肠道类器官结构形成率和出芽数量低于正常（P<0.05）。结论本研究成功建立小鼠小肠类器官培养技术，DCA损伤小肠类器官并影响其生长，长期去除DCA后可部分恢复其功能。 |
| Abstract_FL | Objective To establish a culture system for mouse intestinal organoids and investigate the effect of deoxycholic acid (DCA) on organoids growth. Methods The terminal ileum was collected from 8-month-old C57BL/6 mice. The tissue blocks were treated with EDTA and the crypts were collected and embedded in Matrigel Matrix. Orgnoids growth and buddings were observed in the control group, anhydrous alcohol group, short-term (2 days) 100 μmol/L DCA treatment group, and long-term (10 days) 10 μmol/L DCA treatment group; the orgnoids were further cultured for 10 days after removal of DCA from the medium and observed for orgnoids growth and buddings. Results Short-term treatment with high-concentration DCA resulted in significantly reduced enterosphere formation, enteroids formation, progression from enterospheres to enteroids and number of crypt buds per enteroid (P<0.05), which remained unchanged even after removal of DCA for a short time (P<0.05);long after DCA removal, the enteroids formation rate and number of the crypt buds still remained lower than those in normal organoids (P<0.05). Short-term treatment with low-concentration DCA only resulted in reduced enteroids formation rate and number of crypt buds (P<0.05), and prolonged treatment caused reduced enterospheres formation rate, enteroids formation rate and number of crypt buds (P<0.05). After DCA removal, enteroids formation rate and the number of crypt buds still remained lower than those in the normal group (P<0.05). Conclusion We successfully established an organoids culture system. The presence of DCA in the culture system affects the growth of the organoids, which can partly recover following a prolonged period after the removal of DCA. |
| Author | 楼俪泓 曾悦 周慧 陆颖影 王兴鹏 |
| AuthorAffiliation | 南京医科大学附属上海一院临床医学院消化科，上海200080 上海交通大学附属第一人民医院国际医疗保健中心,上海200080 上海交通大学附属第一人民医院消化科,上海200080 上海市胰腺疾病重点实验室,上海200080 |
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| Author_FL | WANG Xingpeng LOU Lihong LU Yingying ZHOU Hui ZENG Yue |
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| DocumentTitleAlternate | Effect of deoxycholic acid intervention on growth of ileum organoids derived from C57BL/6 mice |
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| Keywords | 类器官 organoids intestinal stem cells 脱氧胆酸 小肠干细胞 deoxycholic acid crypts 隐窝 |
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| Notes | 44-1627/R Objective To establish a culture system for mouse intestinal organoids and investigate the effect of deoxycholic acid （DCA） on organoids growth. Methods The terminal ileum was collected from 8-month-old C57BL/6 mice. The tissue blocks were treated with EDTA and the crypts were collected and embedded in Matrigel Matrix. Orgnoids growth and buddings were observed in the control group, anhydrous alcohol group, short-term （2 days） 100 μmol/L DCA treatment group, and long-term （10 days） 10 μmol/L DCA treatment group; the orgnoids were further cultured for 10 days after removal of DCA from the medium and observed for orgnoids growth and buddings. Results Short-term treatment with high-concentration DCA resulted in significantly reduced enterosphere formation, enteroids formation, progression from enterospheres to enteroids and number of crypt buds per enteroid （P〈0.05）, which remained unchanged even after removal of DCA for a short time （P〈0.05）;long after DCA removal, the enteroids formation rate and numb |
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| Snippet | 目的：建立小鼠小肠类器官培养系统，探讨脱氧胆酸（DCA）对小肠类器官生长的影响。方法取8周龄C57BL/6小鼠末段回肠，EDTA法分离，收集隐窝后Matrigel基质胶包埋，于完整培养基中培养。以无水酒精和正常小肠类器官为对照，予100μmol/L高浓度DCA短期干预2 d，10μmol/L DCA长期干预10... |
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| SubjectTerms | 小肠干细胞 类器官 脱氧胆酸 隐窝 |
| Title | 脱氧胆酸对C57BL/6小鼠回肠类器官生长的影响 |
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