Ovarian cancer is a highly aggressive malignant disease in gynecologic cancer. It is an urgent task to develop three-dimensional (3D) cell models in vitro and dissect the cell progression-related drug resistance mechanisms in vivo. In the present study, RADA16-I peptide has the reticulated nanofiber scaffold networks in hydrogel, which is utilized to develop robust 3D cell culture of a high metastatic human ovarian cancer HO-8910PM cell line accompanied with the counterparts of Matrigel and collagen I. Consequently, HO-8910PM cells were successfully cultivated in three types of hydrogel biomaterials, such as RADA16-I hydrogel, Matrigel, and collagen I, according to 3D cell culture protocols. Designer RADA16-I peptide had well-defined nanofiber networks architecture in hydrogel, which provided nanofiber cell microenvironments analogous to Matrigel and collagen I. 3D-cultured HO-8910PM cells in RADA16-I hydrogel, Matrigel, and collagen I showed viable cell proliferation, proper cell growth, and diverse cell shapes in morphology at the desired time points. For a long 3D cell culture period, HO-8910PM cells showed distinct cell aggregate growth patterns in RADA16-I hydrogel, Matrigel, and collagen I, such as cell aggregates, cell colonies, cell clusters, cell strips, and multicellular tumor spheroids (MCTS). The cell distribution and alignment were described vigorously. Moreover, the molecular expression of integrin β1, E-cadherin and N-cadherin were quantitatively analyzed in 3D-cultured MCTS of HO-8910PM cells by immunohistochemistry and western blotting assays. The chemosensitivity assay for clinical drug responses in 3D context indicated that HO-8910PM cells in three types of hydrogels showed significantly higher chemoresistance to cisplatin and paclitaxel compared to 2D flat cell culture, including IC50 values and inhibition rates. Based on these results, RADA16-I hydrogel is a highly competent, high-profile, and proactive nanofiber scaffold to maintain viable cell proliferation and high cell vitality in 3D cell models, which may be particularly utilized to develop useful clinical drug screening platform in vitro.

中文翻译：

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三种水凝胶生物材料的纳米纤维微环境中高度转移性人类卵巢癌HO-8910PM细胞的三维培养和临床药物反应。

卵巢癌是妇科癌症中高度侵袭性的恶性疾病。体外开发三维（3D）细胞模型并在体内解剖与细胞进程相关的耐药机制是一项紧迫的任务。在本研究中，RADA16-I肽在水凝胶中具有网状纳米纤维支架网络，可用于开发高转移性人卵巢癌HO-8910PM细胞系以及基质胶和胶原蛋白I对应物的稳健3D细胞培养。根据3D细胞培养方案，HO-8910PM细胞已成功在三种类型的水凝胶生物材料中培养，例如RADA16-1水凝胶，基质胶和胶原蛋白I。设计师RADA16-I肽在水凝胶中具有定义明确的纳米纤维网络架构，它提供了类似于Matrigel和I型胶原的纳米纤维细胞微环境。在RADA16-I水凝胶，Matrigel和I型胶原中进行3D培养的HO-8910PM细胞在所需的时间点显示了可行的细胞增殖，适当的细胞生长以及形态多样的细胞形状。在漫长的3D细胞培养期间，HO-8910PM细胞在RADA16-I水凝胶，基质胶和胶原蛋白I中表现出不同的细胞聚集体生长模式，例如细胞聚集体，细胞集落，细胞簇，细胞条和多细胞肿瘤球体（MCTS） ）。强烈描述了细胞分布和排列。此外，通过免疫组织化学和蛋白质印迹分析，在HO-8910PM细胞的3D培养的MCTS中定量分析了整联蛋白β1，E-钙粘着蛋白和N-钙粘着蛋白的分子表达。在3D环境中对临床药物反应的化学敏感性分析表明，与2D平板细胞培养相比，三种类型的水凝胶中的HO-8910PM细胞对顺铂和紫杉醇的化学耐药性显着更高，包括IC50值和抑制率。基于这些结果，RADA16-I水凝胶是一种能胜任，高姿态且积极主动的纳米纤维支架，可在3D细胞模型中维持活细胞增殖和高细胞活力，可特别用于开发有用的体外临床药物筛选平台。