气道上皮干细胞的更新和分化:克服临床级组织工程的挑战性步骤。

IF 7.1 2区 医学 Q1 CELL & TISSUE ENGINEERING
Davide Adamo, Vincenzo Giuseppe Genna, Giulia Galaverni, Chiara Chiavelli, Alessia Merra, Fabio Lepore, Federica Boraldi, Daniela Quaglino, Jessica Evangelista, Filippo Lococo, Graziella Pellegrini
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引用次数: 0

摘要

背景:尽管有挽救生命的潜力,组织工程方法治疗广泛的气管和支气管缺陷仍然面临显著的局限性。一个主要的挑战是在移植生物工程移植物后,无法再生含有长期组织更新所需的适量干细胞的功能性气道上皮。在这种情况下,需要广泛的细胞培养表征,验证分析和质量控制来指导再生过程的每一步。方法:干细胞耗竭通常是由于不理想的培养条件,因此我们测试了临床级培养系统的能力,以支持安全有效的体外扩增和分化原代人气管和支气管上皮细胞。单细胞克隆分析揭示气道基底细胞的异质性,了解组织特异性再生和分化机制。在选择的培养条件下,采用功能试验研究再生上皮的创面愈合能力和紧密性。结果:原代气管支气管上皮细胞显示出令人印象深刻的增殖潜力,允许成熟和功能性上皮的再生而不发生永生化事件。在单细胞水平上的分析可以识别具有体外自我更新能力的基底细胞亚群,将它们与瞬时扩增细胞区分开来。该方法进一步明确了细胞分化的层次结构及其与再生和分化潜能的关系。结论:我们的研究结果表明,原代气道上皮细胞培养可以在体外维持干细胞及其分化谱系。在适当和标准化的条件下培养,气道细胞可以安全有效地用于自体组织工程方法。除了为开发新的先进治疗产品提出的验证分析外,本研究还概述了可能的质量控制,以提高治疗成功率,并在未来的临床应用中最大限度地提高患者的安全性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Airway epithelial stem cell renewal and differentiation: overcoming challenging steps towards clinical-grade tissue engineering.

Background: Despite their life-saving potential, tissue engineering approaches for the treatment of extensive tracheal and bronchial defects still face significant limitations. A major challenge is the inability to regenerate a functional airway epithelium containing the appropriate amount of stem cells required for long-term tissue renewal following transplantation of the bioengineered graft. In this scenario, extensive cell culture characterization, validation assays and quality controls are needed to guide each step of the regeneration process.

Methods: Stem cell depletion is often due to suboptimal culture conditions, therefore we tested the ability of a clinical-grade culture system to support the safe and efficient in vitro expansion and differentiation of primary human tracheal and bronchial epithelial cells. Single-cell clonal analysis was used to unravel the heterogeneity of airway basal cells and to understand tissue-specific regeneration and differentiation mechanisms. Functional assays were used to investigate the wound healing ability and tightness of the regenerated epithelium under the selected culture conditions.

Results: Primary tracheobronchial epithelial cells showed an impressive proliferative potential, allowing the regeneration of a mature and functional epithelium without immortalisation events. Analysis at the single cell level allowed the identification of the subpopulation of basal cells endowed with in vitro self-renewal, distinguishing them from transient amplifying cells. This approach has further defined the hierarchy of cellular differentiation and its correlation with regenerative and differentiation potential.

Conclusions: Our results show that primary airway epithelial cell cultures can maintain stem cells together with their differentiation lineages in vitro. Airway cells can be safely and effectively used in autologous tissue engineering approaches when cultured under appropriate and well-standardised conditions. In addition to the validation assays proposed for the development of new advanced therapy products, this study outlines possible quality controls to enhance therapeutic success and maximise patient safety in future clinical applications.

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来源期刊
Stem Cell Research & Therapy
Stem Cell Research & Therapy CELL BIOLOGY-MEDICINE, RESEARCH & EXPERIMENTAL
CiteScore
13.20
自引率
8.00%
发文量
525
审稿时长
1 months
期刊介绍: Stem Cell Research & Therapy serves as a leading platform for translational research in stem cell therapies. This international, peer-reviewed journal publishes high-quality open-access research articles, with a focus on basic, translational, and clinical research in stem cell therapeutics and regenerative therapies. Coverage includes animal models and clinical trials. Additionally, the journal offers reviews, viewpoints, commentaries, and reports.
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