Ike Soichi, Ueno Koji, Yanagihara Masashi, Mizoguchi Takahiro, Harada Takasuke, Suehiro Kotaro, Kurazumi Hiroshi, Suzuki Ryo, Kondo Tomoko, Murata Tomoaki, Shirasawa Bungo, Morikage Noriyasu, Hamano Kimikazu
Department of Surgery and Clinical Sciences, Yamaguchi University Graduate School of Medicine Ube, Japan.
Department of Molecular Pathology, Yamaguchi University Graduate School of Medicine Ube, Japan.
Am J Transl Res. 2022 Jun 15;14(6):3879-3892. eCollection 2022.
The purpose of this study was to investigate the therapeutic effect of cryopreserved allogenic fibroblast cell sheets in a mouse model of skin ulcers. It is necessary to reduce the cost of regenerative medicine for it to be widely used. We consider that cell sheets could be applied to various diseases if cryopreservation of allogenic cell sheets was possible. In this study, fibroblasts were frozen using a three-dimensional freezer. Freeze-thawed fibroblasts had ~80% cell viability, secreted ≥ 50% vascular endothelial growth factor, hepatocyte growth factor, and stromal derived factor-1α compared with non-frozen fibroblast sheets, and secreted approximately the same amount of transforming growth factor-β1. There was no difference in wound-healing rates in the skin ulcer model between non-frozen and freeze-thawed fibroblast sheets regardless of autologous and allogenic cells. The degree of angiogenesis was comparable between autologous and allogenic cells. The number of CD3-positive cells in healed tissues was larger for allogenic fibroblast sheets compared with autologous fibroblast sheets. However, histopathological images showed that the fibrosis, microvascular density, and healing phase of the wound in allogenic freeze-thawed fibroblast sheets were more similar to autologous freeze-thawed fibroblast sheets than to allogenic non-frozen fibroblast sheets. These results suggest that allogenic freeze-thawed fibroblast sheets may be a promising therapeutic option for refractory skin ulcers.
本研究的目的是在皮肤溃疡小鼠模型中研究冷冻保存的同种异体成纤维细胞片的治疗效果。为了使再生医学得到广泛应用，降低其成本是必要的。我们认为，如果同种异体细胞片的冷冻保存可行，那么细胞片可应用于各种疾病。在本研究中，使用三维冷冻仪对成纤维细胞进行冷冻。与未冷冻的成纤维细胞片相比，冻融后的成纤维细胞具有约80%的细胞活力，分泌的血管内皮生长因子、肝细胞生长因子和基质衍生因子-1α≥50%，且转化生长因子-β1的分泌量大致相同。无论自体和同种异体细胞如何，在皮肤溃疡模型中，未冷冻和冻融后的成纤维细胞片的伤口愈合率没有差异。自体和同种异体细胞之间的血管生成程度相当。与自体成纤维细胞片相比，同种异体成纤维细胞片愈合组织中CD3阳性细胞的数量更多。然而，组织病理学图像显示，同种异体冻融后成纤维细胞片的纤维化、微血管密度和伤口愈合阶段与自体冻融后成纤维细胞片更相似，而与同种异体未冷冻成纤维细胞片不同。这些结果表明，同种异体冻融后成纤维细胞片可能是治疗难治性皮肤溃疡的一种有前景的治疗选择。
