人胚胎干细胞源角质形成细胞用于药物毒理检测的探讨

doi:10.19439/j.sjos.2020.03.006

上海口腔医学 ›› 2020, Vol. 29 ›› Issue (3): 257-261.doi: 10.19439/j.sjos.2020.03.006

人胚胎干细胞源角质形成细胞用于药物毒理检测的探讨

任玉兰¹, 路璐², 李盛林², 俞光岩², 刘鹤³

1.北京大学口腔医学院·口腔医院第一门诊部, 2.口腔颌面外科,3.儿童口腔科,北京 100081

收稿日期:2019-02-13 修回日期:2019-03-27 出版日期:2020-06-25 发布日期:2020-07-29
通讯作者: 刘鹤,E-mail: heliu69@126.com
作者简介:任玉兰（1987-）,女,硕士,住院医师,E-mail:loveonthewalk@163.com
基金资助:
国家国际科技合作专项项目基金（2011DFA32190）

Application of keratinocytes derived from human embryonic stem cells for toxicological evaluation of drugs

REN Yu-lan¹, LU Lu², LI Sheng-lin², YU Guang-yan², LIU He³

1. First Clinical Division, 2. Department of Oral and Maxillofacial Surgery, 3. Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology. Beijing 100081, China

Received:2019-02-13 Revised:2019-03-27 Online:2020-06-25 Published:2020-07-29

摘要/Abstract

摘要： 目的探讨人胚胎干细胞源角质形成细胞（keratinocytes derived from human embryonic stem cells, K-hESCs）用于药物细胞毒理检测反应的可行性,为建立新的生物安全性评价模型提供依据。方法应用CCK-8法检测细胞活性和细胞毒性。观察维A酸（RA）、5氟尿嘧啶（5-FU）、地塞米松（DEX）和青霉素G（PG）对K-hESCs、人原代牙龈上皮细胞（human gingival epithelial cells, HGECs）和人永生化口腔上皮细胞系（human immortalized oral epithelial cells, HIOEC）的细胞毒性反应。采用SPSS 16.0软件包对数据进行统计学分析。结果 RA作用于HGECs、HIOEC和K-hESCs 3种角质形成细胞后,其药物半数抑制浓度（IC₅₀）分别为6.1±0.03、5.62±0.05和6.58±0.02;5-FU作用于3种细胞后,IC₅₀分别为1.65±0.02、3.00±0.02和1.72±0.04;DEX作用于3种细胞后,IC₅₀分别为113.67±0.014、328±0.002和126.17±0.05;PG作用于3种细胞后,IC₅₀分别为2200±1.34、3795±2.42和2880±1.5。4种药物对HIOEC和HGECs的IC₅₀有统计学差异（P<0.05）。对K-hESCs和HIOEC的IC₅₀差异也有统计学意义（P<0.05）,但对K-hESCs和HGECs的IC₅₀差异无统计学意义（P>0.05）。结论在一般细胞毒性上, K-hESCs的药物半数抑制浓度比HIOEC更接近HGECs,可模拟人体正常细胞的反应。

关键词: 人胚胎干细胞, 角质形成细胞, 细胞毒性

Abstract: PURPOSE: To preliminarily explore the cytotoxicological responses of keratinocytes derived from human embryonic stem cells(K-hESCs) to drugs, and to provide a basis for the establishment of a new biosafety evaluation model. METHODS: CCK-8 assay was used to detect cell viability and cytotoxicity. The detection of pharmacological response was observed and compared when K-hESCs directly derived from human embryonic stem cells, human gingival epithelial cells (HGECs), and human immortalized oral epithelial cells (HIOECs) were treated with retinoic acid (RA), 5-fluorouracil(5-FU), dexamethasone(DEX), and penicillin G(PG). Statistical analysis was performed using SPSS 16.0 software package. RESULTS: After drugs were applied to HGECs, HIOEC and K-hESCs, the half inhibitory concentrations (IC₅₀) of RA was 6.1±0.03, 5.62±0.05 and 6.58±0.02, respectively. The IC₅₀ of 5-FU was 1.65±0.02,3.00±0.02 and 1.72±0.04, respectively. The IC₅₀ of DEX was 113.67±0.014,328±0.002 and 126.17±0.05, respectively. The IC₅₀ of PG was 2200±1.34,3795±2.42 and 2880±1.5, respectively. The IC₅₀ of the four drugs between HIOEC and HGECs had significant differences(P<0.05), but there was no significant difference in IC₅₀ between K-hESCs and HGECs(P>0.05). The IC₅₀ of K-hESCs and HIOEC also had significant differences(P<0.05). CONCLUSIONS: IC₅₀ of K-hESCs was closer to HGECs than HIOEC. It was speculated that K-hESCs could simulate the response of normal human cells in cytotoxicity study.

Key words: Human embryonic stem cells, Keratinocytes, Cytotoxicity

中图分类号:

R739.8

任玉兰, 路璐, 李盛林, 俞光岩, 刘鹤. 人胚胎干细胞源角质形成细胞用于药物毒理检测的探讨[J]. 上海口腔医学, 2020, 29(3): 257-261.

REN Yu-lan, LU Lu, LI Sheng-lin, YU Guang-yan, LIU He. Application of keratinocytes derived from human embryonic stem cells for toxicological evaluation of drugs[J]. Shanghai Journal of Stomatology, 2020, 29(3): 257-261.

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人胚胎干细胞源角质形成细胞用于药物毒理检测的探讨

Application of keratinocytes derived from human embryonic stem cells for toxicological evaluation of drugs

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