Wang Xuanzhi, Dai Xingliang, Zhang Xinzhi, Li Xinda, Xu Tao, Lan Qing
Department of Neurosurgery, The Second Affiliated Hospital of Soochow University, Suzhou, 215004, People's Republic of China.
Medprin Biotech GmbH, Gutleutstraße 163-167, Frankfurt am Main, D-60327, Germany; Department of Mechanical Engineering, Biomanufacturing Center, Tsinghua University, Beijing, 100084, People's Republic of China.
Biochem Biophys Res Commun. 2018 Apr 15;498(4):1052-1057. doi: 10.1016/j.bbrc.2018.03.114. Epub 2018 Mar 17.
Cancer stem cells (CSCs), being tumor-initiating with self-renewal capacity and heterogeneity, are most likely the cause of tumor resistance, reoccurrence and metastasis. To further investigate the role of CSCs in tumor biology, there is a need to develop an effective culture system to grow, maintain and enrich CSCs. Three-dimensional (3D) cell culture model has been widely used in tumor research and drug screening. Recently, researchers have begun to utilize 3D models to culture cancer cells for CSCs enrichment. In this study, glioma cell line was cultured with 3D porous chitosan (CS) scaffolds or chitosan-hyaluronic acid (CS-HA) scaffolds to explore the possibility of glioma stem cells (GSCs)-like cells enrichment, to study the morphology, gene expression, and in vivo tumorigenicity of 3D scaffolds cells, and to compare results to 2D controls. Results showed that glioma cells on both CS and CS-HA scaffolds could form tumor cell spheroids and increased the expression of GSCs biomarkers compared to conventional 2D monolayers. Furthermore, cells in CS-HA scaffolds had higher expression levels of epithelial-to-mesenchymal transition (EMT)-related gene. Specifically, the in vivo tumorigenicity capability of CS-HA scaffold cultured cells was greater than 2D cells or CS scaffold cultured cells. It is indicated that the chemical composition of scaffold plays an important role in the enrichment of CSCs. Our results suggest that CS-HA scaffolds have a better capability to enrich GSCs-like cells and can serve as a simple and effective way to cultivate and enrich CSCs in vitro to support the study of CSCs biology and development of novel anti-cancer therapies.
癌症干细胞（CSCs）具有肿瘤起始、自我更新能力和异质性，很可能是肿瘤耐药、复发和转移的原因。为了进一步研究CSCs在肿瘤生物学中的作用，需要开发一种有效的培养系统来培养、维持和富集CSCs。三维（3D）细胞培养模型已广泛应用于肿瘤研究和药物筛选。最近，研究人员开始利用3D模型培养癌细胞以富集CSCs。在本研究中，用3D多孔壳聚糖（CS）支架或壳聚糖-透明质酸（CS-HA）支架培养胶质瘤细胞系，以探索富集胶质瘤干细胞（GSCs）样细胞的可能性，研究3D支架细胞的形态、基因表达和体内致瘤性，并将结果与二维对照进行比较。结果表明，与传统的二维单层细胞相比，CS和CS-HA支架上的胶质瘤细胞均可形成肿瘤细胞球体，并增加了GSCs生物标志物的表达。此外，CS-HA支架中的细胞上皮-间质转化（EMT）相关基因的表达水平更高。具体而言，CS-HA支架培养的细胞的体内致瘤能力大于二维培养的细胞或CS支架培养的细胞。这表明支架的化学成分在CSCs的富集中起着重要作用。我们的结果表明，CS-HA支架具有更好的富集GSCs样细胞的能力，可作为一种简单有效的体外培养和富集CSCs的方法，以支持CSCs生物学研究和新型抗癌疗法的开发。
