酸刺激对人体细胞特性影响的研究

许庆庆; 段甦; 翟所强; 张罗

doi:10.13201/j.issn.1001-1781.2019.07.005

酸刺激对人体细胞特性影响的研究

- 首都医科大学附属北京同仁医院耳鼻咽喉科(北京, 100853)

详细信息

通讯作者: 张罗,E-mail:dr.luozhang@139.com

中图分类号: R764.3

收稿日期: 2019-03-10

Experimental study on the effect of acid stimulation on human cell characteristics

- Department of Otorhinolaryngology, Beijing Tongren Hospital, Capital Medical University, Beijing, 100853, China

More Information

Corresponding author: ZHANG Luo

Received Date: 10 March 2019

摘要

摘要: 目的:探索体外培养的体细胞经过低pH处理后的特性和分化特征,为感音神经性聋提供新的治疗策略。方法:选取人成熟体细胞作为目标细胞,选择几种pH值不同溶液分别处理细胞,对细胞的形态进行观察,从碱性磷酸酶(AKP)活性、免疫组织化学染色、分子生物学等方面对细胞的特性进行鉴定,选出最适应pH值。对诱导后细胞体外进行神经干定向诱导,再将基膜以胆红素处理,建立感音神经性聋体外模型,验证诱导后细胞的体外分化能力以及治疗功能;验证方法为形态学、AKP活性、免疫组织化学、分子生物学等手段。结果:实验组细胞生长明显优于对照组;实验组AKP活性高于对照组;Nanog和Oct4的表达在2组均为阳性;将处理后的细胞转移到神经干细胞培养液中,Nestin表达阳性。结论:低pH处理后的人体细胞显示出与诱导多能干细胞相似的特性,但其功能和治疗效果有待进一步研究。
- 低pH值 /
- 形态学 /
- 碱性磷酸酶 /
- 免疫荧光
Abstract: Objective: The purpose of the present study was to explore the characteristics and differentiation of somatic cells in vitro undergoing a low pH treatment, so as to provide new therapeutic strategies for treating sensorineural hearing loss. Method: The human mature somatic cells were selected as the target cells, and the cells were treated with different pH values to observe the cell morphology. The cell characteristics were identified from alkaline phosphatase (AKP) activity, immunohistochemical staining and molecular biology, and the most suitable pH value was selected. In addition, a mouse model of the cochlear lesion was constructed using bilirubin. Subsequently, the characteristics and therapeutic effect of somatic cells undergoing low pH treatment were examined by morphology, AKP activity, immunofluorescence assay and Q-PCR.Result: The cell growth of the experimental group was significantly better than those in the control group. The activity of AKP in the experimental group was higher than that in the control group. The expression of Nanog and Oct4 was both positive in the two groups. When the cells were changed to neurobasol medium, the marker of Nestin was positive.Conclusion: The human somatic cells undergoing a low pH treatment showed the similar characteristics as those of induced pluripotent stem (iPS) cells; although the functions and therapeutic effect of these altered human somatic cells need to be further studied.
- low pH /
- morphology /
- alkaline phosphatase /
- immunofluorescence assay

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