a Department of Pharmacology, School of Medicine , Shahid Beheshti University of Medical Sciences , Tehran , Iran.
b Department of Biomaterials, Faculty of Biomedical Engineering , Amirkabir University of Technology , Tehran , Iran.
Artif Cells Nanomed Biotechnol. 2018;46(sup1):876-884. doi: 10.1080/21691401.2018.1438452. Epub 2018 Feb 23.
Placenta-derived amniotic epithelial cells (AECs), a great cell source for tissue engineering and stem cell therapy, are immunologically inert in their native state; however, immunological changes in these cells after culture and differentiation have challenged their applications. The aim of this study was to investigate the effect of 2D and 3D scaffolds on human lymphocyte antigens (HLA) expression by AECs. The effect of different preparation parameters including pre-freezing time and temperature was evaluated on 3D chitosan-gelatine scaffolds properties. Evaluation of MHC class I, HLA-DR and HLA-G expression in AECs after 7 d culture on 2D bed and 3D scaffold of chitosan-gelatine showed that culture of AECs on the 2D substrate up-regulated MHC class I and HLA-DR protein markers on AECs surface and down-regulated HLA-G protein. In contrast, 3D scaffold did not increase protein expression of MHC class I and HLA-DR. Moreover, HLA-G protein expression remained unchanged in 3D culture. These results confirm that 3D scaffold can remain AECs in their native immunological state and modification of physical properties of the scaffold is a key regulator of immunological markers at the gene and protein expression levels; a strategy which circumvents rejection challenge of amniotic stem cells to be translated into the clinic.
胎盘来源的羊膜上皮细胞（AECs）是组织工程和干细胞治疗的极佳细胞来源，在其天然状态下具有免疫惰性；然而，这些细胞在培养和分化后的免疫变化挑战了它们的应用。本研究旨在探讨 2D 和 3D 支架对 AECs 人类淋巴细胞抗原（HLA）表达的影响。评估了不同的制备参数对 3D 壳聚糖-明胶支架性能的影响，包括预冻时间和温度。7 d 后，在 2D 床和 3D 壳聚糖-明胶支架上培养 AECs，评估 AECs 中 MHC Ⅰ类、HLA-DR 和 HLA-G 的表达，结果表明 AECs 在 2D 基质上培养会上调 AECs 表面 MHC Ⅰ类和 HLA-DR 蛋白标志物，并下调 HLA-G 蛋白。相比之下，3D 支架不会增加 MHC Ⅰ类和 HLA-DR 的蛋白表达。此外，3D 培养中 HLA-G 蛋白表达保持不变。这些结果证实，3D 支架可以使 AECs 保持其天然免疫状态，并且支架物理性质的改变是基因和蛋白表达水平上免疫标志物的关键调节剂；这种策略可以避免羊膜干细胞的排斥挑战，从而转化为临床应用。
