Department of Electrical, Computer, and Biomedical Engineering, Union College, Schenectady, NY, USA.
Cryobiology. 2022 Apr;105:41-49. doi: 10.1016/j.cryobiol.2021.12.003. Epub 2021 Dec 16.
Despite progress in many aspects of controlling cell behavior within synthetic three-dimensional hydrogels, approaches to cryopreserve these systems - encompassing the protection of both encapsulated cell viability and network bioactive functions - are lacking. Here, we demonstrate the retention of encapsulated human mesenchymal stromal cell (hMSC) viability following in situ cryopreservation regardless of cell line, material system, or storage duration. Further, the preservation extends to network bioactive functions, with hMSCs cryopreserved within degradable and adhesive hyaluronic-acid (HA) based hydrogels exhibiting degradation-mediated spreading within the gels equivalent to their non-frozen counterparts. Finally, the platform cryopreservation protocol preserves multi-lineage cellular differentiation capacity, with encapsulated hMSCs in non-degradable and adhesive/degradable HA-based hydrogels undergoing rates of adipogenesis and osteogenesis, respectively, equivalent to those in non-frozen gels on a per-cell basis. Collectively, these findings indicate a versatile platform technology that contributes to an increased understanding of three-dimensional cell-matrix interactions, and which may enable the indefinite cryopreservation of tissue engineering constructs for clinical applications.
尽管在控制合成三维水凝胶中细胞行为方面取得了许多进展，但缺乏冷冻保存这些系统的方法 - 包括保护封装细胞活力和网络生物活性功能。在这里，我们证明了无论细胞系、材料系统或储存时间如何，原位冷冻保存后封装的人骨髓间充质基质细胞（hMSC）活力得以保留。此外，这种保存扩展到网络生物活性功能，在可降解和粘附透明质酸（HA）基水凝胶中冷冻保存的 hMSC 表现出与非冷冻对应物相当的凝胶内降解介导的扩散。最后，该平台冷冻保存方案保留了多谱系细胞分化能力，在非降解和粘附/可降解 HA 基水凝胶中封装的 hMSC 经历脂肪生成和成骨的速率，分别相当于非冷冻凝胶中的每细胞基础。总的来说，这些发现表明这是一种通用的平台技术，有助于增加对三维细胞基质相互作用的理解，并且可能能够无限期地冷冻保存组织工程构建体用于临床应用。
