Ren Yu-Lan, Lu Lu, Li Sheng-Lin, Yu Guang-Yan, Liu He
First Clinical Division, Peking University School and Hospital of Stomatology. Beijing 100081, China. E-mail:
Shanghai Kou Qiang Yi Xue. 2020 Jun;29(3):257-261.
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.
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.
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).
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.
初步探究人胚胎干细胞来源的角质形成细胞(K-hESCs)对药物的细胞毒理学反应,为建立新的生物安全性评价模型提供依据。
采用CCK-8法检测细胞活力和细胞毒性。观察并比较维甲酸(RA)、5-氟尿嘧啶(5-FU)、地塞米松(DEX)和青霉素G(PG)作用于直接来源于人胚胎干细胞的K-hESCs、人牙龈上皮细胞(HGECs)和人永生化口腔上皮细胞(HIOECs)时的药理反应。使用SPSS 16.0软件包进行统计分析。
将药物作用于HGECs、HIOEC和K-hESCs后,RA的半数抑制浓度(IC50)分别为6.1±0.03、5.62±0.05和6.58±0.02。5-FU的IC50分别为1.65±0.02、3.00±0.02和1.72±0.04。DEX的IC50分别为113.67±0.014、328±0.002和126.17±0.05。PG的IC50分别为2200±1.34、3795±2.42和2880±1.5。四种药物在HIOEC和HGECs之间的IC50有显著差异(P<0.05),但K-hESCs与HGECs之间的IC50无显著差异(P>0.05)。K-hESCs与HIOEC的IC50也有显著差异(P<0.05)。
K-hESCs的IC50比HIOEC更接近HGECs。推测在细胞毒性研究中K-hESCs可模拟正常人细胞的反应。