Medical School of Chinese PLA, Beijing, China.
Department of Stomatology, The First Medical Center, Chinese PLA General Hospital, Beijing, China.
PeerJ. 2024 Oct 31;12:e18418. doi: 10.7717/peerj.18418. eCollection 2024.
Epidermal stem cells (EpSCs) are crucial for wound healing and tissue regeneration, and traditional culture methods often lead to their inactivation. It is urgent to increase the yield of high quality EpSCs. In this study, primary EpSCs were isolated and cultured in a serum-free, feeder-free culture system. EpSCs are then expanded in a dynamic 3D environment using a rotating cell culture system (RCCS) with biodegradable porous microcarriers (MC). Over a period of 14 days, the cells self-assembled into microtissues with superior cell proliferation compared to 3D static culture. Immunofluorescence and qPCR analyses consistently showed that the stemness of the 3D microtissues was preserved, especially the COL17A1 associated with anti-aging was highly expressed in RCCS induced microtissues. experiments demonstrated that the group treated with 3D microtissues loaded with EpSCs showed enhanced early wound healing, and the injectable 3D microtissues were more conducive to maintaining cell viability and differentiation potential. Our study provides valuable insights into the dynamic 3D culture of EpSCs and introduces an injectable therapy using 3D microtissues loaded with EpSCs, which provides a new and effective approach for cell delivery and offering a promising strategy for guiding the regeneration of full-thickness skin defects.
表皮干细胞（EpSCs）对于伤口愈合和组织再生至关重要，而传统的培养方法往往会导致其失活。因此，迫切需要提高高质量 EpSCs 的产量。在本研究中，我们从原发性 EpSCs 中分离并在无血清、无饲养细胞的培养体系中进行培养。然后，我们使用具有生物可降解多孔微载体（MC）的旋转细胞培养系统（RCCS）在动态 3D 环境中对 EpSCs 进行扩增。在 14 天的时间内，细胞自我组装成微组织，与 3D 静态培养相比，细胞增殖能力得到了显著提高。免疫荧光和 qPCR 分析一致表明，3D 微组织保留了干细胞特性，特别是与抗衰老相关的 COL17A1 在 RCCS 诱导的微组织中高度表达。实验表明，用 EpSCs 负载的 3D 微组织处理的组表现出增强的早期伤口愈合能力，并且可注射的 3D 微组织更有利于维持细胞活力和分化潜能。我们的研究为 EpSCs 的动态 3D 培养提供了有价值的见解，并介绍了一种使用 EpSCs 负载的 3D 微组织的可注射治疗方法，为细胞递送提供了一种新的有效方法，并为指导全层皮肤缺损的再生提供了有前途的策略。
