Saberianpour Shirin, Rezaie Nezhad Zamani Arezoo, Karimi Abbas, Ahmadi Mahdi, Khatami Neda, Pouyafar Ayda, Rahbarghazi Reza, Nouri Mohammad
Department of Molecular Medicine, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Adv Pharm Bull. 2020 Jan;10(1):141-145. doi: 10.15171/apb.2020.019. Epub 2019 Dec 11.
Today, there is an urgent need to develop a three-dimentional culture systems mimicking native condition in order to screen potency of drugs and possibly any genetic alterations in tumor cells. Due to the existence of limitations in animal models, the development of three dimensional systems is highly recommended. To this end, we encapsulated human colon adenocarcinoma cell line HT29 with alginate-poly-L-lysine (Alg-PLL) microspheres and the rate of epithelial-mesenchymal transition was monitored. Cells were randomly divided into three groups; control, alginate and Alg-PLL. To encapsulate cells, we mixed HT-29 cells (1 × 10 ) with 1 mL of 0.05% PLL and 1% Alg mixture and electrosprayed into CaCl2 solution by using a high-voltage power. Cells from all groups were maintained at 37˚C in a humidified atmosphere containing 5% CO for 7 days. Cell viability was assessed by MTT assay. To monitor the stemness feature, we measured the transcription of genes such as , and by using real-time PCR analysis. Addition of PLL to Alg in a hallowed state increased the cell survival rate compared to the control and Alg groups (<0.05). Cells inside Alg-PLL tended to form microcellular aggregates while in Alg microspheres an even distribution of HT-29 cells was found. Real-time PCR analysis showed the up-regulation of , and in Alg-PLL microspheres compared to the other groups, showing the acquisition of stemness feature (<0.05). This study showed that hallow Alg-PLL microspheres increased the epithelialmesenchymal transition rate after 7 days in condition. Such approaches could be touted as appropriate models for drug screening.
如今,迫切需要开发一种模拟天然条件的三维培养系统,以便筛选药物的效力以及肿瘤细胞中可能存在的任何基因改变。由于动物模型存在局限性,强烈建议开发三维系统。为此,我们用藻酸盐-聚-L-赖氨酸(Alg-PLL)微球包裹人结肠腺癌细胞系HT29,并监测上皮-间质转化的速率。细胞被随机分为三组:对照组、藻酸盐组和Alg-PLL组。为了包裹细胞,我们将HT-29细胞(1×10)与1 mL 0.05%的PLL和1%的Alg混合物混合,并使用高压电源将其电喷雾到CaCl2溶液中。所有组的细胞在含有5% CO的湿润气氛中于37˚C下培养7天。通过MTT法评估细胞活力。为了监测干性特征,我们使用实时PCR分析测量了诸如等基因的转录。与对照组和Alg组相比,在中空状态下向Alg中添加PLL可提高细胞存活率(<0.05)。Alg-PLL内的细胞倾向于形成微细胞聚集体,而在Alg微球中发现HT-29细胞分布均匀。实时PCR分析表明,与其他组相比,Alg-PLL微球中的、和上调,表明获得了干性特征(<0.05)。这项研究表明,中空的Alg-PLL微球在条件下7天后可提高上皮-间质转化速率。此类方法可被视为适用于药物筛选的模型。