Charbonneau André M, Tran Simon D
McGill Craniofacial Tissue Engineering and Stem Cells Laboratory, Faculty of Dentistry, McGill University, Montréal, QC H3A 2B2, Canada.
Materials (Basel). 2020 Oct 28;13(21):4807. doi: 10.3390/ma13214807.
The egg yolk plasma (EYP)-a translucent fraction of the egg yolk (EY) obtained by centrifugation-was tested as a developmentally encouraging, cost-effective, biomaterial for salivary gland (SG) tissue engineering. To find optimal incubating conditions for both the human NS-SV-AC SG acinar cell line and SG fibroblasts, cells were stained with Live/Dead. The cellular contents of 96-well plates were analyzed by high content screening image analysis. Characteristically, the EYP biomaterial had lipid and protein content resembling the EY. On its own, the EYP was non-conducive to cell survival. EYP's pH of 6 mainly contributed to cell death. This was demonstrated by titrating EYP's pH with different concentrations of either commercial cell culture media, NaOH, or egg white (EW). These additives improved SG mesenchymal and epithelial cell survival. The best combinations were EYP diluted with (1) 70% commercial medium, (2) 0.02 M NaOH, or (3) 50% EW. Importantly, commercial medium-free growth was obtained with EYP + NaOH or EYP + EW. Furthermore, 3D cultures were obtained as a result of EW's gelatinous properties. Here, the isolation, characterization, and optimization of three EYP-based biomaterial combinations are shown; two were free of commercial medium or supplements and supported both SG cells' survival.
蛋黄血浆（EYP）是通过离心获得的蛋黄（EY）的半透明部分，作为一种对唾液腺（SG）组织工程有促进发育作用且经济高效的生物材料进行了测试。为了找到适合人类NS-SV-AC SG腺泡细胞系和SG成纤维细胞的最佳培养条件，细胞用活/死细胞染色试剂盒进行染色。通过高内涵筛选图像分析对96孔板中的细胞内容物进行分析。典型地，EYP生物材料的脂质和蛋白质含量与EY相似。就其本身而言，EYP不利于细胞存活。EYP的pH值为6主要导致细胞死亡。用不同浓度的商业细胞培养基、氢氧化钠或蛋清（EW）滴定EYP的pH值证明了这一点。这些添加剂提高了SG间充质细胞和上皮细胞的存活率。最佳组合是用（1）70%商业培养基、（2）0.02 M氢氧化钠或（3）50% EW稀释的EYP。重要的是，使用EYP + 氢氧化钠或EYP + EW可实现无商业培养基培养。此外，由于EW的凝胶特性获得了三维培养物。在此展示了三种基于EYP的生物材料组合的分离、表征和优化；其中两种不含商业培养基或补充剂，并支持两种SG细胞的存活。
