Automated procedure for biomimetic de-cellularized lung scaffold supporting alveolar epithelial transdifferentiation
Automated procedure for biomimetic de-cellularized lung scaffold supporting alveolar epithelial transdifferentiation
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DOI：
10.1016/j.biomaterials.2013.09.055
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发表时间：
2013-12-01
期刊：
影响因子：
14
通讯作者：
Finck, Christine M.
中科院分区：
文献类型：
--
作者：
Girard, Eric D.;Jensen, Todd J.;Finck, Christine M.
制备无细胞和细胞碎片、免疫惰性、保留必要的细胞外基质（ECM）的去细胞化肺支架的最佳方法尚未确定。在此，我们比较了使用恒压（CP）或恒流量（CF）的基于洗涤剂的自动脱细胞方法，以及之前发表的使用手动压力（MP）注入和冲洗脱细胞剂的方案。评估每种方法产生的去细胞化肺是否存在剩余的ECM蛋白和免疫刺激物质，如核酸和细胞内物质。我们的研究结果表明，CP和MP方法更有效地去除细胞物质，但保留ECM蛋白的差异。CP方法具有更快、可重复的去细胞过程的附加好处。为了评估去细胞化支架维持上皮细胞的功能能力，气管内接种表达C10肺泡上皮细胞（AEC）的GFP。值得注意的是，CP去细胞化的肺能够支持C10-GFP细胞从ii型样表型到i型样表型的生长和自发分化。(C) 2013 Elsevier Ltd.版权所有。
The optimal method for creating a de-cellularized lung scaffold that is devoid of cells and cell debris, immunologically inert, and retains necessary extracellular matrix (ECM) has yet to be identified. Herein, we compare automated detergent-based de-cellularization approaches utilizing either constant pressure (CP) or constant flow (CF), to previously published protocols utilizing manual pressure (MP) to instill and rinse out the de-cellularization agents. De-cellularized lungs resulting from each method were evaluated for presence of remaining ECM proteins and immunostimulatory material such as nucleic acids and intracellular material. Our results demonstrate that the CP and MP approaches more effectively remove cellular materials but differentially retain ECM proteins. The CP method has the added benefit of being a faster, reproducible de-cellularization process. To assess the functional ability of the de-cellularized scaffolds to maintain epithelial cells, intra-tracheal inoculation with GFP expressing C10 alveolar epithelial cells (AEC) was performed. Notably, the CP de-cellularized lungs were able to support growth and spontaneous differentiation of C10-GFP cells from a type II-like phenotype to a type I-like phenotype. (C) 2013 Elsevier Ltd. All rights reserved.