Laboratory of Cell Culture, Plastic Surgery Division, Universidade Federal de São Paulo, São Paulo, Brazil. vanina
Int Wound J. 2010 Oct;7(5):339-48. doi: 10.1111/j.1742-481X.2010.00698.x.
In vitro studies with keloid fibroblasts frequently present contradictory results. This may occur because keloids present distinct genotypic and phenotypic characteristics in its different regions, such as the peripheral region in relation to the central region. We suggest an explant model for keloid fibroblasts harvesting, standardising the initial processing of keloid samples to obtain fragments from different regions, considering its biological differences, for primary cell culture. The different keloid regions were delimited and fragments were obtained using a 3-mm diameter punch. To remove fragments from the periphery, the punch was placed in one longitudinal line extremity, respecting the lesion borders. For the central region, it was placed in the intersection of lines at the level of the largest longitudinal and transversal axes, the other fragments being removed centrifugally in relation to the first one. Primary fibroblast culture was carried out by explant. Flow cytometry analysis showed cell cycle differences between the groups, confirming its different origins and biological characteristics. In conclusion, our proposed model proved itself efficient for keloid fibroblast isolation from specific regions and cultivation. Its simplicity and ease of execution may turn it into an important tool for studying the characteristics of the different keloid-derived fibroblasts in culture.
体外研究瘢痕成纤维细胞常呈现出相互矛盾的结果。这可能是因为瘢痕在不同区域表现出明显的基因型和表型特征，例如与中央区域相比，周边区域。我们建议采用瘢痕成纤维细胞采集的离体模型，标准化瘢痕样本的初始处理，以获得不同区域的片段，考虑到其生物学差异，用于原代细胞培养。用 3 毫米直径的打孔器划定不同的瘢痕区域并获取片段。为了从周边去除碎片，打孔器放在一条纵向线的末端，以符合病变边界。对于中央区域，将其放置在最大纵轴和横轴的交点处，其余的碎片以相对于第一个碎片的离心方式去除。原代纤维母细胞培养通过离体进行。流式细胞术分析显示各组细胞周期存在差异，证实其起源和生物学特性不同。总之，我们提出的模型被证明可有效分离特定区域的瘢痕成纤维细胞并进行培养。其简单易用的特点可能使其成为研究不同瘢痕衍生纤维母细胞在培养中的特征的重要工具。
