Fu Jingjing, Chen Feng, Chai Huihui, Gao Lixia, Lv Xiaohui, Yu Ling
School of Preclinical Medicine, North Sichuan Medical College, Nanchong 637000, China.
Institute for Clean Energy & Advanced Materials, School of Materials & Energy, Southwest University, Chongqing 400715, China.
Nanomaterials (Basel). 2022 Feb 28;12(5):808. doi: 10.3390/nano12050808.
A gelatin@non-woven fabric (gelatin@NWF) hybrid scaffold with tailored micropore structures was fabricated by lyophilizing, using gelatin to support cells and the NWF matrix as a frame to enforce the mechanical stability of gelatin. By freezing the gelatin and NWF hybrid in liquid nitrogen and subsequently lyophilizing and crosslinking the process, the gelatin@NWF scaffold was prepared to support cell growth and promote cell aggregation and spheroids' formation. The results indicated that by tuning the lyophilizing temperature, the micropore size on the gelatin could be tailored. Consequently, tumor spheroids can be formed on gelatin@NWF scaffolds with honeycomb-like pores around 10 µm. The cell spheroids formed on the tailored gelatin@NWF scaffold were characterized in cancer stem cell (CSC)-associated gene expression, chemotherapy drug sensitivity, and motility. It was found that the expression of the CSC-associated biomarkers SOX2, OCT4, and ALDH1A1 in gene and protein levels in DU 145 cell spheres formed on gelatin@NWF scaffolds were significantly higher than in those cells grown as monolayers. Moreover, cells isolated from spheroids grown on gelatin@NWF scaffold showed higher drug resistance and motility. Tumor spheroids can be formed on a long-term storage scaffold, highlighting the potential of gelatin@NWF as a ready-to-use scaffold for tumor cell sphere generation and culturing.
通过冻干法制备了具有定制微孔结构的明胶@无纺布（gelatin@NWF）复合支架，利用明胶来支持细胞生长，并以无纺布基质作为框架来增强明胶的机械稳定性。通过将明胶和NWF复合物在液氮中冷冻，随后进行冻干和交联处理，制备出gelatin@NWF支架以支持细胞生长并促进细胞聚集和球体形成。结果表明，通过调节冻干温度，可以定制明胶上的微孔尺寸。因此，在具有约10 µm蜂窝状孔的gelatin@NWF支架上可以形成肿瘤球体。对在定制的gelatin@NWF支架上形成的细胞球体进行了癌症干细胞（CSC）相关基因表达、化疗药物敏感性和运动性的表征。研究发现，在gelatin@NWF支架上形成的DU 145细胞球体中，CSC相关生物标志物SOX2、OCT4和ALDH1A1在基因和蛋白质水平上的表达明显高于单层生长的细胞。此外，从在gelatin@NWF支架上生长的球体中分离出的细胞表现出更高的耐药性和运动性。肿瘤球体可以在长期储存的支架上形成，这突出了gelatin@NWF作为一种现成的用于肿瘤细胞球体生成和培养的支架的潜力。
