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张程程,彭艳,陈海交,杨建文,刘薇薇,刘丽.成年SD大鼠胃窦Cajal间质细胞分离、培养及鉴定[J].湖南中医药大学学报英文版,2017,37(11):1226-1230.[Click to copy
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| 成年SD大鼠胃窦Cajal间质细胞分离、培养及鉴定 |
| 张程程,彭艳,陈海交,杨建文,刘薇薇,刘丽 |
| (湖南中医药大学针灸推拿学院针灸生物信息分析重点实验室, 湖南 长沙 410208) |
| 摘要: |
| 目的 探讨成年SD大鼠胃窦起搏细胞Cajal间质细胞（interstitial cells of cajal，ICC）的原代分离、培养及鉴定方法，为深入研究其生理功能提供条件。方法 8~9周龄SD成年大鼠，禁食24 h，乙醚吸入麻醉，颈椎脱臼处死。无菌条件下采用精细解剖方法快速取出胃窦平滑肌组织，将其剪成1~2 mm3小块后，使用Ⅱ型胶原酶消化法于37℃培养箱内消化60 min，离心后过200目筛，接种于DMEM/F12完全培养基（含5 ng/mL干细胞因子）中进行培养。用酪氨酸蛋白激酶受体c-kit特异性抗体免疫荧光染色鉴定细胞类型。结果 培养72 h后，于倒置显微镜下观察，可见细胞贴壁，呈不规则三角形、星形或梭形，有多个突起；随着培养时间的延长，各突起彼此相互连接形成网络；此原代培养细胞c-kit抗体免疫荧光染色呈阳性。结论 成功建立了一套由成年SD大鼠胃窦组织采用Ⅱ型胶原酶消化法原代培养ICC的方法，为ICC的生理功能、病理改变以及胃肠道生理与病理关系的研究奠定了基础。 |
| 关键词: 胃窦 Cajal间质细胞 细胞培养技术 |
| DOI：10.3969/j.issn.1674-070X.2017.11.014 |
| Received:November 24, 2016 |
| 基金项目:国家自然科学基金项目（81403487）；湖南省教育厅青年基金（14B128）。 |
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| Isolation, Culture and Identification of Interstitial Cells of Cajal in Gastric Antrum of Adult Rats |
| ZHANG Chengcheng,PENG Yan,CHEN Haijiao,YANG Jianwen,LIU Weiwei,LIU Li |
| (Key Acu-Moxibustion Laboratory of Biological Information Analysis, Institute of Acupuncture, Moxibustion and Massage, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective In order to provide the conditions for further study of its physiological function, the method of isolation, culture and identification of interstitial cells of Cajal (ICC) in the gastric antrum of adult SD rats was investigated. Methods SD rats at 8-9 weeks, were anesthetized with ether and killed by cervical dislocation. The specimens were cut into 1-2mm3 small pieces, digested with collagenase type Ⅱ in a 37℃ incubator for 60 min, centrifuged and passed through a 200-mesh sieve. The tissues were inoculated in DMEM/F12 complete medium (containing 5 ng/mL stem cell factor). Cell type was identified by immunofluorescence staining using tyrosine protein kinase receptor c-kit specific antibody. Results After culture for 72 h, the cells adhered to the wall under the inverted microscope, which were irregular triangular, star-shaped, or spindle-shaped, with multiple protrusions. With the prolongation of culture period, the protrusions were connected to each other to form a network. The cells were positive for immunofluorescence staining of c-kit antibody. Conclusion This study established a method for primary culture of ICC from type Ⅱ collagenase digestion of gastric antral smooth muscle in adult SD rats. It laid a foundation for the study of the relationship between ICC and gastrointestinal tract in physiological function and pathologic changes. |
| Key words: gastric antrum interstitial cells of Cajal cell culture techniques |
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