| 摘要: |
| 近年来，水凝胶在细胞和类器官培养和冷冻保存方面表现出显著优势，尤其在细胞冷冻保存方面。在该研究中，选用喷雾法，以三价铁和钙为交联剂，采用低成本且易获得的喷枪制备了海藻酸钙和海藻酸铁水凝胶，并用该方法包封了HEK293T细胞和HepG2细胞，研究了不同交联液对包封细胞时细胞存活率的影响，发现交联时间越长对细胞损伤越大，与海藻酸钠溶液交联后发现凝胶后降低了细胞在交联液中的损伤。还制备了不同质量浓度海藻酸钠溶液（1%、1.5%、3%）与CaCl2（0.2 mol/L）交联的水凝胶，包封HEK293T细胞并培养7 d，发现细胞均可以很好地在水凝胶中培养。最后，研究了包封后细胞冷冻保存的存活率，结果表明：HEK293T细胞和HepG2细胞在两种体积浓度的二甲基亚砜（DMSO）为保护剂的前提下存活率均显著高于未包封组，包封前HEK293T细胞的存活率为33.16% ± 2.70%（10% DMSO）和16.75% ± 2.3%（5% DMSO），而包封后的细胞存活率增至76.51% ± 5.32%（10% DMSO）和60.86% ± 2.41%（5% DMSO）。对于HepG2细胞，细胞存活率则从包封前的48.93% ± 3.06%（10% DMSO）和36.22% ± 2.54%（5% DMSO）增至包封后的78.79% ± 4.43%（10% DMSO）和64.64% ± 3.13%（5% DMSO）。在乙二醇（1 mol/L）+丙二醇（1.5 mol/L）+海藻糖（1 mol/L）作为保护剂的前提下，玻璃化保存的HEK293T细胞的存活率由包封前的33.5% ± 0.8%增至79% ± 3.76%。数据表明，冷冻保存后包封细胞的存活率明显高于未包封细胞，离子交联海藻酸盐水凝胶可以在细胞冷冻保存中保护细胞。 |
| 关键词: 海藻酸盐 水凝胶 细胞 玻璃化 冷冻保存 |
| DOI： |
| 投稿时间：2023-02-21 修订日期：2023-03-19
录用日期：2023-04-03 |
| 基金项目:国家自然科学基金（51776130）资助项目。 |
|
| Examining Ionic Cross-Linked Alginate Hydrogels in Cell Culture and Cryopreservation |
| Li Qiye,Wang Shige,Liu Baolin |
| (Institute of Biothermal and Technology, University of Shanghai for Science and Technology;School of Materials and Chemistry, University of Shanghai for Science and Technology) |
| Abstract: |
| In recent years, hydrogels have exhibited considerable advantages in cell and organoids culture as well as cryopreservation, particularly in cell cryopreservation. In this study, the spray method was selected, and ferric iron and calcium were used as cross-linking agents. The hydrogels were prepared using a low-cost and readily available airbrush. Subsequently, calcium alginate and iron alginate hydrogels were prepared, and HEK293T cells and HepG2 cells were encapsulated using this method. The study also investigated the impact of cross-linking solutions on cell viability during encapsulation, revealing that longer crosslinking times led to greater cell damage. Additionally, it was observed that crosslinking with sodium alginate solution reduced cell damage in the crosslinking solution. Furthermore, hydrogels crosslinked with CaCl2 (0.2 mol/L) using sodium alginate solution at various mass concentrations (1%, 1.5%, and 3%) were prepared. HEK293T cells were encapsulated and cultured for 7 d under these conditions, and successful cell culture was observed across all concentrations. Finally, the viability of cell cryopreservation after encapsulation was examined. The results showed that the viability of HEK293T and HepG2 cells, under the precondition of using two different volumetric concentrations of dimethyl sulfoxide as protective agents, was significantly higher than that of the unencapsulated group. Specifically, for HEK293T cells before encapsulation, cell viability was 33.16% ± 2.70% (10% DMSO) and 16.75% ± 2.3% (5% DMSO). After encapsulation, cell viability increased to 76.51% ± 5.32% (10% DMSO) and 60.86% ± 2.41% (5% DMSO). Similarly, for HepG2 cells, cell viability increased from 48.93% ± 3.06% (10% DMSO) and 36.22% ± 2.54% (5% DMSO) to 78.79% ± 4.43% (10% DMSO) and 64.64% ± 3.13% (5% DMSO) after encapsulation. Moreover, when using ethylene glycol (1 mol/L) + propylene glycol (1.5 mol/L) + trehalose (1 mol/L) as a protective agent, the viability of HEK293T cells after vitrification increased from 33.5% ± 0.8% to 79% ± 3.76% after encapsulation. These results strongly indicate that encapsulated cells exhibit significantly higher viability after cryopreservation compared to unencapsulated cells, emphasizing the protective capacity of ionic cross-linked alginate hydrogels during cryopreservation. |
| Key words: alginate hydrogel cell vitrification cryopreservation |
