Department of General Pathology, Pomeranian Medical University, Powstanców Wlkp. 72, 70-111 Szczecin, Poland.
Department of Histology and Embryology, Pomeranian Medical University, Powstanców Wlkp. 72, 70-111 Szczecin, Poland.
Int J Med Sci. 2021 Feb 3;18(6):1519-1531. doi: 10.7150/ijms.46265. eCollection 2021.
Chronological skin ageing is an inevitable physiological process that results in thin and sagging skin, fine wrinkles, and gradual dermal atrophy. The main therapeutic approaches to soft tissue augmentation involve using dermal fillers, where natural fillers, such as autologous fibroblasts, are involved in generating dermal matrix proteins. The aim of this study was to determine the global transcriptome profile of three passages of dermal autologous fibroblasts from a male volunteer, focusing on the processes of the cell cycle and cell proliferation status to estimate the optimal passage of the tested cells with respect to their reimplantation. We performed K-means clustering and validation of the expression of the selected mRNA by qRT-PCR. Ten genes were selected (ANLN, BUB1, CDC20, CCNA2, DLGAP5, MKI67, PLK1, PRC1, SPAG5, and TPX2) from the top five processes annotated to cluster 5. Detailed microarray analysis of the fibroblast genes indicated that the cell population of the third passage exhibited the highest number of upregulated genes involved in the cell cycle and cell proliferation. In all cases, the results of qRT-PCR confirmed the differences in expression of the selected mRNAs between fibroblasts from the primary culture (C0) and from the first (C1), second (C2), and third (C3) cell passage. Our results thus suggest that these cells might be useful for increasing fibroblast numbers after reimplantation into a recipient's skin, and the method used in this study seems to be an excellent tool for autologous transplantation allowing the rejuvenation of aging skin.
皮肤的老化是一种不可避免的生理过程，它会导致皮肤变薄、松弛、出现细小皱纹，并逐渐出现真皮萎缩。软组织填充是治疗皮肤老化的主要方法之一，其中包括使用真皮填充剂，这些填充剂涉及到利用自体成纤维细胞来产生真皮基质蛋白。本研究的目的是确定一名男性志愿者的 3 代自体真皮成纤维细胞的全转录组谱，重点研究细胞周期和细胞增殖状态的过程，以评估用于细胞再植入的测试细胞的最佳传代数。我们进行了 K-means 聚类分析，并通过 qRT-PCR 对选定的 mRNA 表达进行了验证。从注释为聚类 5 的前 5 个过程中选择了 10 个基因（ANLN、BUB1、CDC20、CCNA2、DLGAP5、MKI67、PLK1、PRC1、SPAG5 和 TPX2）。对成纤维细胞基因的详细微阵列分析表明，第 3 代细胞群体中涉及细胞周期和细胞增殖的上调基因数量最多。在所有情况下，qRT-PCR 的结果都证实了原代培养（C0）和成纤维细胞第 1 代（C1）、第 2 代（C2）和第 3 代（C3）传代之间选定 mRNA 的表达差异。因此，我们的研究结果表明，这些细胞在重新植入受体皮肤后可能有助于增加成纤维细胞的数量，并且本研究中使用的方法似乎是一种用于自体移植的优秀工具，可用于皮肤老化的年轻化。
