Department of Molecular Science and Technology, Ajou University, Suwon, Korea.
Cell Therapy Center, Ajou Medical Center, Suwon, 16499, Korea.
Cell Tissue Res. 2022 Aug;389(2):289-308. doi: 10.1007/s00441-022-03638-5. Epub 2022 May 28.
Stem cells are known to have excellent regenerative ability, which is primarily facilitated by indirect paracrine factors, rather than via direct cell replacement. The regenerative process is mediated by the release of extracellular matrix molecules, cytokines, and growth factors, which are also present in the media during cultivation. Herein, we aimed to demonstrate the functionality of key factors and mechanisms in skin regeneration through the analysis of conditioned media derived from fetal stem cells. A series of processes, including 3D pellet cultures, filtration and lyophilization is developed to fabricate human fetal cartilage-derived progenitor cells-conditioned media (hFCPCs-CM) and its useful properties are compared with those of human bone marrow-derived MSCs-conditioned media (hBMSCs-CM) in terms of biochemical characterization, and in vitro studies of fibroblast behavior, macrophage polarization, and burn wound healing. The hFCPCs-CM show to be devoid of cellular components but to contain large amounts of total protein, collagen, glycosaminoglycans, and growth factors, including IGFBP-2, IGFBP-6, HGF, VEGF, TGF β3, and M-CSF, and contain a specific protein, collagen alpha-1(XIV) compare with hBMSCs-CM. The therapeutic potential of hFCPCs-CM observes to be better than that of hBMSCs-CM in the viability, proliferation, and migration of fibroblasts, and M2 macrophage polarization in vitro, and efficient acceleration of wound healing and minimization of scar formation in third-degree burn wounds in a rat model. The current study shows the potential therapeutic effect of hFCPCs and provides a rationale for using the secretome released from fetal progenitor cells to promote the regeneration of skin tissues, both quantitatively and qualitatively. The ready-to-use product of human fetal cartilage-derived progenitor cells-conditioned media (hFCPCs-CM) are fabricated via a series of techniques, including a 3D culture of hFCPCs, filtration using a 3.5 kDa cutoff dialysis membrane, and lyophilization of the CM. hFCPCs-CM contains many ECM molecules and biomolecules that improves wound healing through efficient acceleration of M2 macrophage polarization and reduction of scar formation.
干细胞具有出色的再生能力，主要是通过间接旁分泌因子来实现，而不是通过直接细胞替代。再生过程是通过细胞外基质分子、细胞因子和生长因子的释放来介导的，这些分子和因子在培养过程中也存在于培养基中。在此，我们旨在通过分析来源于胎儿干细胞的条件培养基来证明皮肤再生中关键因子和机制的功能。我们开发了一系列工艺，包括 3D 微球培养、过滤和冻干，以制备人胎儿软骨源祖细胞条件培养基（hFCPCs-CM），并比较其与人间充质干细胞条件培养基（hBMSCs-CM）在生化特性、成纤维细胞行为、巨噬细胞极化和烧伤创面愈合的体外研究中的特性。hFCPCs-CM 不含细胞成分，但含有大量总蛋白、胶原蛋白、糖胺聚糖和生长因子，包括 IGFBP-2、IGFBP-6、HGF、VEGF、TGFβ3 和 M-CSF，并含有一种特定的蛋白质，胶原蛋白 alpha-1(XIV)，与 hBMSCs-CM 相比。hFCPCs-CM 的治疗潜力优于 hBMSCs-CM，表现在成纤维细胞的活力、增殖和迁移，以及体外 M2 巨噬细胞极化，以及在大鼠三度烧伤模型中加速创面愈合和减少瘢痕形成。本研究显示了 hFCPCs 的潜在治疗效果，并为利用胎儿祖细胞分泌的细胞外基质来促进皮肤组织的再生提供了依据，无论是在数量上还是质量上。通过一系列技术，包括 hFCPCs 的 3D 培养、使用 3.5 kDa 截止透析膜过滤和 CM 的冻干，制备了即用型人胎儿软骨源祖细胞条件培养基（hFCPCs-CM）。hFCPCs-CM 含有许多细胞外基质分子和生物分子，通过有效促进 M2 巨噬细胞极化和减少瘢痕形成来加速伤口愈合。
