Kim Min Ho, Wu Wen Hao, Choi Jee Hyun, Kim Jihyun, Jun Jin Hyun, Ko Yong, Lee Jong Hun
Eulji Medi-Bio Research Institute, Eulji University, Seoul, Republic of Korea.
Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul, Republic of Korea.
Wound Repair Regen. 2018 Dec;26 Suppl 1:S9-S18. doi: 10.1111/wrr.12579. Epub 2017 Nov 8.
Keratinocytes and fibroblasts cells play important roles in the skin-wound healing process and are the cell types activated by trauma. Activated cells participate in epithelialization, granulation, scar tissue formation, wound remodeling, and angiogenesis via a series of cellular activities including migration and proliferation. Previous studies reported that the conditioned medium (CM) of adipose-derived stem cells (ADSCs) stimulated the migration and proliferation of cell types involved in the skin wound healing process; however, these studies only show ADSC-CM effects that were obtained using 2-dimensional (2D) culture. Recently, 3-dimensional (3D) culture has been considered as a more physiologically appropriate system than 2D culture for ADSC cultures; therefore, ADSC-CM was collected from 3D culture (ADSC-CM-3D) and compared with ADSC-CM from 2D culture (ADSC-CM-2D) to investigate the effects on the migration and proliferation of human keratinocytes (HaCaTs) and fibroblasts. The migrations of the HaCaT cells and fibroblasts were significantly higher with ADSC-CM-3D compared with the 2D culture; similarly, the proliferation of HaCaT cells was also highly stimulated by ADSC-CM-3D. Proteomic analyses of the ADSC-CM revealed that collagens and actins were highly expressed in the 3D-culture system. Chitinase 3-like 1 (CHI3L1), tissue inhibitor of metalloproteinases (TIMP), and galectin-1 were specifically expressed only in ADSC-CM-3D. Especially, through antibody neutralization, galectin-1 in ADSC-CM-3D was found to be an important factor for the migration of human keratinocytes. Therefore, these results suggest that ADSC-CM-3D was more effective in the wound healing than ADSC-CM-2D, and galectin-1 in ADSC-CM-3D was could be a promising option for skin-wound healing. Furthermore, the differential expressions of several ADSC-CM proteins between the 2D- and 3D-culture systems may be used as basic information for the development of efficient wound-healing strategies.
角质形成细胞和成纤维细胞在皮肤伤口愈合过程中发挥着重要作用，是受创伤激活的细胞类型。活化的细胞通过包括迁移和增殖在内的一系列细胞活动参与上皮形成、肉芽形成、瘢痕组织形成、伤口重塑和血管生成。先前的研究报道，脂肪来源干细胞（ADSCs）的条件培养基（CM）可刺激参与皮肤伤口愈合过程的细胞类型的迁移和增殖；然而，这些研究仅显示了使用二维（2D）培养获得的ADSC-CM的作用。最近，三维（3D）培养被认为是一种比2D培养更适合ADSC培养的生理系统；因此，从3D培养中收集ADSC-CM（ADSC-CM-3D）并与2D培养中的ADSC-CM（ADSC-CM-2D）进行比较，以研究其对人角质形成细胞（HaCaTs）和成纤维细胞迁移和增殖的影响。与2D培养相比，ADSC-CM-3D处理的HaCaT细胞和成纤维细胞的迁移明显更高；同样，ADSC-CM-3D也高度刺激了HaCaT细胞的增殖。对ADSC-CM的蛋白质组学分析表明，胶原蛋白和肌动蛋白在3D培养系统中高表达。几丁质酶3样1（CHI3L1）、金属蛋白酶组织抑制剂（TIMP）和半乳糖凝集素-1仅在ADSC-CM-3D中特异性表达。特别是，通过抗体中和发现，ADSC-CM-3D中的半乳糖凝集素-1是人类角质形成细胞迁移的重要因素。因此，这些结果表明，ADSC-CM-3D在伤口愈合方面比ADSC-CM-2D更有效，并且ADSC-CM-3D中的半乳糖凝集素-1可能是皮肤伤口愈合的一个有前景的选择。此外，2D和3D培养系统之间几种ADSC-CM蛋白的差异表达可作为开发高效伤口愈合策略的基础信息。
