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首页> 外文期刊>Journal of biomedical materials research. Part B, Applied biomaterials. >Characterization of viability and proliferation of alginate-poly-L-lysine-alginate encapsulated myoblasts using flow cytometry.
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Characterization of viability and proliferation of alginate-poly-L-lysine-alginate encapsulated myoblasts using flow cytometry.

机译：使用流式细胞仪表征藻酸盐-聚-L-赖氨酸-藻酸盐包裹的成肌细胞的活力和增殖。

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Genetically modified cells encapsulated in alginate-poly-L-lysine-alginate (APA) are being developed to deliver therapeutic products to treat a variety of diseases. The characterization of the encapsulated cells thus becomes paramount. This study reports a novel method to assess the viability, granularity and proliferation of encapsulated cells based on flow cytometry. The in vitro viability of encapsulated G8 murine myoblasts secreting canine FVIII (cFVIII) measured by flow cytometry was comparable to the traditional trypan blue exclusion method and both correlated with cFVIII secretion levels. In contrast, after implantation into mice, only viability measured by flow cytometry correlated with cFVIII secretion. Further, flow cytometry analysis of encapsulated cells maintained in vitro and in vivo revealed a greater fraction of granular cells compared to free cells, suggesting that encapsulation influences the morphology (cytoplasmic composition) of cells within APA microcapsules. Interestingly, the proliferation study showed that encapsulated cells proliferate faster, on average, and were more heterogeneous in vivo compared to in vitro culture conditions, suggesting that encapsulated cell proliferation is complex and environment-dependent. In conclusion, we show that flow cytometry analysis allows for a more consistent and comprehensive examination of encapsulated cells to aid in the development of cell therapy protocols.
机译：正在开发封装在藻酸盐-聚-L-赖氨酸-藻酸盐（APA）中的转基因细胞，以提供治疗多种疾病的治疗产品。因此，封装细胞的表征至关重要。这项研究报告了一种基于流式细胞术评估封装细胞的活力，粒度和增殖的新方法。通过流式细胞仪测量的封装的分泌犬FVIII（cFVIII）的G8鼠成肌细胞的体外生存力与传统的锥虫蓝排除法相当，并且都与cFVIII分泌水平相关。相反，植入小鼠后，仅通过流式细胞术测量的生存能力与cFVIII分泌相关。此外，在体外和体内维持的封装细胞的流式细胞仪分析显示，与游离细胞相比，粒状细胞的比例更高，这表明封装影响了APA微胶囊内细胞的形态（细胞质组成）。有趣的是，增殖研究表明，与体外培养条件相比，封装的细胞平均在体内增殖更快，并且在体内的异质性更高，这表明封装的细胞增殖是复杂的并且是环境依赖性的。总之，我们表明流式细胞仪分析可以对封装的细胞进行更一致和更全面的检查，以帮助开发细胞治疗方案。

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