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Living biointerfaces based on non-pathogenic bacteria support stem cell differentiation

机译:基于非致病细菌的活体生物界面支持干细胞分化

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摘要

Lactococcus lactis, a non-pathogenic bacteria, has been genetically engineered to express the III7–10 fragment of human fibronectin as a membrane protein. The engineered L. lactis is able to develop biofilms on different surfaces (such as glass and synthetic polymers) and serves as a long-term substrate for mammalian cell culture, specifically human mesenchymal stem cells (hMSC). This system constitutes a living interface between biomaterials and stem cells. The engineered biofilms remain stable and viable for up to 28 days while the expressed fibronectin fragment induces hMSC adhesion. We have optimised conditions to allow long-term mammalian cell culture, and found that the biofilm is functionally equivalent to a fibronectin-coated surface in terms of osteoblastic differentiation using bone morphogenetic protein 2 (BMP-2) added to the medium. This living bacteria interface holds promise as a dynamic substrate for stem cell differentiation that can be further engineered to express other biochemical cues to control hMSC differentiation.
机译:乳酸乳球菌(一种非致病性细菌)经过基因工程改造,可将人纤连蛋白的III7-10片段表达为膜蛋白。经过改造的乳酸乳球菌能够在不同的表面(例如玻璃和合成聚合物)上形成生物膜,并可以作为哺乳动物细胞培养的长期底物,特别是人间充质干细胞(hMSC)。该系统构成了生物材料和干细胞之间的活泼界面。在表达的纤连蛋白片段诱导hMSC粘附的同时,经过工程改造的生物膜可保持稳定并存活长达28天。我们优化了条件,以允许长期哺乳动物细胞培养,并发现在成骨细胞分化方面,使用添加到培养基中的骨形态发生蛋白2(BMP-2),生物膜在功能上等同于纤连蛋白包被的表面。这种活细菌的界面有望成为干细胞分化的动态底物,可以进一步工程化以表达其他生化线索来控制hMSC的分化。

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