Characterization of in vitro model based on primary culture of human mammary epithelial cells (hMECs).

This data set contains results related to the characterization of an appropriate in vitro model based on human mammary epithelial cells (hMECs). The objective of the research was to verify the accuracy of hMECs as solid translational model for the study of mammary epithelial barrier. The results showed the characterization (morphological data; epithelial markers Epithelial-Cadherin (E-Cad) and Cytokeratin 18 (CK18); gene expression of drugs transporters and doubling Time (DT)), maintaining of hMEC primary culture. The hMECs maintained until P6 showed a typical cobblestone morphology and doubling time data showed a mean value of 36.6 ± 3.25 hours (see data file CONCEPTION_WP3_hMECs_DoublingTime_20240723.csv). The hMEC expressed epithelial markers Epithelial-Cadherin (E-Cad) and Cytokeratin 18 (CK18), confirming their epithelial origin (see data file CONCEPTION_WP3_hMECs_FlowCytometer_20240723.csv). In addition we analyze the gene expression of 84 drugs transporters (by using a commercial kit, RT2) and the Ct value was reported in CONCEPTION_WP3_hMECs_RT2ARRAY_20240723.xlsx) The transcriptional profile of drug transporters showed a differential level of gene expression ranging from ΔCt values (Ct mean reference genes – Ct interest gene) very negative (lower expression) to ΔCt values less negative (higher expression). Only 3 were not detectable (SLC22A9, SCCO1B3 and SLCO1B3). Collectively, the results show that primary culture of hMECs express epithelial cell markers and tight junctions molecule and express the main drug transporters, suggesting that it will be used to study epithelial barrier functions.

本数据集包含基于人乳腺上皮细胞（human mammary epithelial cells, hMECs）构建的适宜体外模型的表征相关研究结果。本研究的核心目标为验证hMECs作为乳腺上皮屏障研究的稳健转化模型的准确性。研究结果涵盖原代hMECs维持培养过程中的各项表征数据，包括形态学资料、上皮标志物上皮型钙黏蛋白（Epithelial-Cadherin, E-Cad）与细胞角蛋白18（Cytokeratin 18, CK18）的表达情况、药物转运蛋白基因表达水平及倍增时间（doubling time, DT）。传代至第6代（P6）的hMECs呈现典型的铺路石样形态，其倍增时间均值为36.6±3.25小时，相关数据详见数据文件CONCEPTION_WP3_hMECs_DoublingTime_20240723.csv。hMECs可表达上皮标志物上皮型钙黏蛋白（E-Cad）与细胞角蛋白18（CK18），证实其上皮细胞起源，相关流式细胞术检测数据详见CONCEPTION_WP3_hMECs_FlowCytometer_20240723.csv。此外，本研究采用商用RT2试剂盒对84种药物转运蛋白的基因表达水平进行了分析，相关Ct值数据详见CONCEPTION_WP3_hMECs_RT2ARRAY_20240723.xlsx。药物转运蛋白的转录谱显示基因表达水平存在显著差异，ΔCt值（参考基因平均Ct值-目的基因Ct值）范围从极负值（对应低表达）到弱负值（对应高表达）不等。其中仅3种基因未被检测到，分别为SLC22A9、SCCO1B3及SLCO1B3。综上，本研究结果表明，原代hMECs可表达上皮细胞标志物及紧密连接分子，并可表达主要的药物转运蛋白，提示该模型可用于乳腺上皮屏障功能的相关研究。