Soon Chin Fhong, Tee Kian Sek, Wong Soon Chuan, Nayan Nafarizal, Ahmad Mohd Khairul, Sefat Farshid, Sultana Naznin, Youseffi Mansour
Biosensor and Bioengineering Lab, MiNT-SRC, Faculty of Electrical and Electronic Engineering, Universiti Tun Hussein Onn Malaysia, 86400, Parit Raja, Batu Pahat, Johor, Malaysia.
Faculty of Engineering and Informatics, Medical and Healthcare Technology Department, University of Bradford, Bradford, BD7 1DP, UK.
Cytotechnology. 2018 Feb;70(1):13-29. doi: 10.1007/s10616-017-0168-2. Epub 2017 Nov 30.
Growing three dimensional (3D) cells is an emerging research in tissue engineering. Biophysical properties of the 3D cells regulate the cells growth, drug diffusion dynamics and gene expressions. Scaffold based or scaffoldless techniques for 3D cell cultures are rarely being compared in terms of the physical features of the microtissues produced. The biophysical properties of the microtissues cultured using scaffold based microencapsulation by flicking and scaffoldless liquid crystal (LC) based techniques were characterized. Flicking technique produced high yield and highly reproducible microtissues of keratinocyte cell lines in alginate microcapsules at approximately 350 ± 12 pieces per culture. However, microtissues grown on the LC substrates yielded at lower quantity of 58 ± 21 pieces per culture. The sizes of the microtissues produced using alginate microcapsules and LC substrates were 250 ± 25 μm and 141 ± 70 μm, respectively. In both techniques, cells remodeled into microtissues via different growth phases and showed good integrity of cells in field-emission scanning microscopy (FE-SEM). Microencapsulation packed the cells in alginate scaffolds of polysaccharides with limited spaces for motility. Whereas, LC substrates allowed the cells to migrate and self-stacking into multilayered structures as revealed by the nuclei stainings. The cells cultured using both techniques were found viable based on the live and dead cell stainings. Stained histological sections showed that both techniques produced cell models that closely replicate the intrinsic physiological conditions. Alginate microcapsulation and LC based techniques produced microtissues containing similar bio-macromolecules but they did not alter the main absorption bands of microtissues as revealed by the Fourier transform infrared spectroscopy. Cell growth, structural organization, morphology and surface structures for 3D microtissues cultured using both techniques appeared to be different and might be suitable for different applications.
培养三维（3D）细胞是组织工程领域一项新兴的研究。3D细胞的生物物理特性可调节细胞生长、药物扩散动力学和基因表达。基于支架或无支架的3D细胞培养技术，很少根据所产生的微组织的物理特征进行比较。对使用基于支架的微囊化轻弹技术和基于无支架液晶（LC）技术培养的微组织的生物物理特性进行了表征。轻弹技术在藻酸盐微囊中产生了高产率且高度可重复的角质形成细胞系微组织，每次培养约350±12个。然而，在LC基质上生长的微组织产量较低，每次培养58±21个。使用藻酸盐微囊和LC基质产生的微组织大小分别为250±25μm和141±70μm。在这两种技术中，细胞通过不同的生长阶段重塑为微组织，并且在场发射扫描显微镜（FE-SEM）中显示出良好的细胞完整性。微囊化将细胞包裹在多糖的藻酸盐支架中，细胞运动空间有限。而LC基质允许细胞迁移并自堆叠成多层结构，细胞核染色显示了这一点。根据活细胞和死细胞染色发现，使用这两种技术培养的细胞都是有活力的。染色的组织学切片表明，这两种技术都产生了紧密复制内在生理条件的细胞模型。藻酸盐微囊化和基于LC的技术产生了含有相似生物大分子的微组织，但傅里叶变换红外光谱显示它们并未改变微组织的主要吸收带。使用这两种技术培养的3D微组织的细胞生长、结构组织、形态和表面结构似乎不同，可能适用于不同的应用。
