Characterization of viability and proliferation of alginate-poly-L-lysine-alginate encapsulated myoblasts using flow cytometry
Characterization of viability and proliferation of alginate-poly-L-lysine-alginate encapsulated myoblasts using flow cytometry
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DOI：
10.1002/jbm.b.31648
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发表时间：
2010-08-01
影响因子：
3.4
通讯作者：
Hortelano, Gonzalo
中科院分区：
文献类型：
--
作者：
Thakur, Ajit;Sengupta, Ruchira;Hortelano, Gonzalo
海藻酸盐-聚l -赖氨酸-海藻酸盐（APA）包裹的基因修饰细胞正在被开发，以提供治疗产品来治疗各种疾病。因此，包膜细胞的表征变得至关重要。本研究报告了一种基于流式细胞术评估被包被细胞活力、粒度和增殖的新方法。用流式细胞术检测包封的分泌犬FVIII （cFVIII）的G8小鼠成肌细胞体外活力与传统的台锥蓝排除法相当，且均与cFVIII分泌水平相关。相比之下，植入小鼠后，只有流式细胞术测量的活力与cFVIII分泌相关。此外，对体外和体内被包被的细胞进行流式细胞术分析发现，与游离细胞相比，颗粒细胞的比例更高，这表明包被影响了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，被包裹细胞在体内的平均增殖速度更快，异质性更强，这表明被包裹细胞的增殖是复杂的，依赖于环境。总之，我们表明，流式细胞术分析允许更一致和全面的检查包膜细胞，以帮助细胞治疗方案的发展。(C) 2010 Wiley期刊公司[J] .中国生物医学工程学报，2010,31(2):444 - 444。
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. (C) 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 94B: 296-304, 2010.