Introduction

Human mesenchymal stem cells (hMSCs) holds great promise for managing several clinical conditions. However, the low engraftment efficiency and obscurity to harvest these cells without compromising the cellular viability, structural and functional properties from the culture niche still remain major obstacles for preparing intact regenerative constructs. Although few studies have demonstrate different methods for generating cell-liberated amniotic scaffolds, a common method for producing completely cell-liberated amnion (D-HAM) and chorion (D-HCM) scaffolds and their cytocompatibility with hMSCs yet to be demonstrated.

Methods

A common process was developed for preparing D-HAM and D-HCM scaffolds for assessing hMSCs engraftment efficiency, proliferation and molecular shifts to generate cell-laden biological discs. The structural and functional integrity of D-HAM and D-HCM was evaluated using different parameters. The compatibility and proliferation efficiency of hMSCs with D-HAM and D-HCM was evaluated.

Results

Histological analysis revealed completely nucleic acid-free D-HAM and D-HCM scaffolds with intact extracellular matrix, mechanical and biological properties almost similar to the native membranes. Human MSCs were able to adhere and engraft on D-HCM better than D-HAM and expanded faster. Ultrastructural observations, crystal violet staining and expression studies showed better structural and functional integrity of hMSCs on D-HCM than D-HAM and control conditions.

Conclusion

A common, simple and reliable process of decellularization can generate large number of cell-liberated amniotic scaffolds in lesser time. D-HCM has better efficiency for hMSCs engraftment and proliferation and can be utilized for preparing suitable cell-laden constructs for tissue engineering applications.

中文翻译：

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脱细胞羊膜和绒毛膜支架的制造以开发生物工程载细胞构建体

介绍

人类间充质干细胞 (hMSCs) 在管理多种临床疾病方面具有很大的前景。然而，在不损害培养生态位的细胞活力、结构和功能特性的情况下收获这些细胞的低植入效率和晦涩难懂仍然是制备完整再生结构的主要障碍。尽管很少有研究证明了产生细胞释放羊膜支架的不同方法，但生产完全释放细胞的羊膜 (D-HAM) 和绒毛膜 (D-HCM) 支架的常用方法及其与 hMSCs 的细胞相容性尚未得到证实。

方法

开发了一种用于制备 D-HAM 和 D-HCM 支架的通用过程，用于评估 hMSCs 的植入效率、增殖和分子转移，以产生载有细胞的生物盘。使用不同的参数评估 D-HAM 和 D-HCM 的结构和功能完整性。评估了 hMSCs 与 D-HAM 和 D-HCM 的相容性和增殖效率。

结果

组织学分析显示完全无核酸的 D-HAM 和 D-HCM 支架具有完整的细胞外基质，机械和生物学特性几乎与天然膜相似。与 D-HAM 相比，人类 MSC 能够更好地粘附和移植到 D-HCM 上，并且扩张速度更快。超微结构观察、结晶紫染色和表达研究表明，hMSC 在 D-HCM 上的结构和功能完整性优于 D-HAM 和对照条件。

结论

一种常见、简单且可靠的脱细胞过程可以在更短的时间内生成大量细胞释放的羊膜支架。D-HCM 对 hMSCs 的植入和增殖具有更高的效率，可用于制备适合组织工程应用的载细胞构建体。