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首页> 美国卫生研究院文献>Regenerative Therapy >Preparation of EpH4 and 3T3L1 cells aggregates incorporating gelatin hydrogel microspheres for a cell condition improvement
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Preparation of EpH4 and 3T3L1 cells aggregates incorporating gelatin hydrogel microspheres for a cell condition improvement

机译：制备含有明胶水凝胶微球的EpH4和3T3L1细胞聚集体以改善细胞状况

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The objective of this study is to prepare three dimensional (3D) of mouse mammary epithelial EpH4 and mouse preadipocyte 3T3L1 cells in the presence of gelatin hydrogel microspheres (GM) and evaluate the effect of GM presence on the survival and functions of cells in the 3D cell aggregates. Gelatin was dehydrothermally crosslinked at 140 °C for 48 h in a water-in-oil emulsion state to obtain the GM with average diameters of 50 and 200 μm, followed by treatment with fibronectin (FN). EpH4 and/or 3T3L1 cells were cultured with or without the FN-treated GM in round U-bottom wells of 96-multiwell culture plates which had been coated with poly (vinyl alcohol) (PVA) to allow the cells to form their aggregates. On the other hand, EpH4 cells were precultured with the FN-treated GM, and then continued to culture with 3T3L1 cells in the same condition described above. The EpH4 cells attached onto the GM in the cell number dependent manner, irrespective of their size. When 3T3L1 cells were incubated with the original and GM-preincubated EpH4 cells in the presence of both the FN-treated GM, the number of alive cells in the aggregates was significantly high compared with that for the absence of FN-treated GM. In addition, higher β-casein expression level of EpH4 cells in EpH4/3T3L1 cells aggregates in the presence of FN-treated GM was observed than that of cells in the absence of FN-treated GM. Laminin secretion was also promoted for the cells aggregates cultured with FN-treated GM. It is concluded that the presence of FN-treated GM in the EpH4/3T3L1 cells aggregates gave a better condition to cells, resulting in an enhanced generation of β-casein from EpH4 cells in the aggregates.
机译：这项研究的目的是在明胶水凝胶微球（GM）存在的情况下制备小鼠乳腺上皮EpH4和小鼠前脂肪细胞3T3L1细胞的三维（3D）并评估GM存在对3D细胞存活和功能的影响细胞聚集体。将明胶在140°C下以油包水乳液状态进行48h脱氢热交联，得到平均直径为50和200μm的GM，然后用纤连蛋白（FN）处理。将EpH4和/或3T3L1细胞在有FN处理的GM或不经过FN处理的GM的情况下，在96孔培养板的U形底部圆形孔中培养，该孔已用聚乙烯醇（PVA）包被，以使细胞形成聚集体。另一方面，将EpH4细胞与经FN处理的GM一起预培养，然后在上述相同条件下继续与3T3L1细胞一起培养。 EpH4细胞以细胞数依赖性方式附着在GM上，而不管其大小如何。当在FN处理的GM均存在的情况下，将3T3L1细胞与原始和GM预孵育的EpH4细胞一起孵育时，与不存在FN处理的GM相比，聚集体中的活细胞数量明显较高。另外，观察到在存在FN处理的GM的情况下，EpH4 / 3T3L1细胞聚集体中的EpH4细胞的β-酪蛋白表达水平比不存在FN处理的GM的细胞高。对于用FN处理的GM培养的细胞聚集体，层粘连蛋白的分泌也被促进。结论是，EpH4 / 3T3L1细胞聚集体中经过FN处理的GM的存在为细胞提供了更好的条件，从而导致聚集体中EpH4细胞中β-酪蛋白的生成增加。

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