He Yan, Zhang Shengpeng, Bao Wu, Xue Yi, Yin Bin, Cheng Xialin, Zhang Zexin, Jia Chiyu
School of Medicine, Xiamen University, Xiamen, China.
Department of Burns and Plastic and Wound Repair Surgery, Xiang'an Hospital of Xiamen University, School of Medicine, Xiamen University, Xiamen, China.
J Cosmet Dermatol. 2022 Dec;21(12):7131-7139. doi: 10.1111/jocd.15416. Epub 2022 Oct 14.
Keloid (KD) is a complex fibroproliferative disease, but the exact mechanisms underlying keloid pathogenesis remain to be elucidated. The primary keloid fibroblasts (KFs) culture in vitro has always been a fundamental measure to study the pathogenesis of keloid. However, the traditional primary culture methods have some limitations, such as a long culture cycle, low specimen utilization rate and so on.
Improve the keloid explants culture method sts.
MATERIALS & METHODS: We proposed an improved new "explants multiple culture method"-reusing keloid explants for primary culture and harvesting the primary KFs in specific culture times. Meanwhile, the purity, proliferation, apoptosis, migration, invasion, extracellular matrix synthesis, and some fibrosis and inflammation-related proteins of KFs obtained from the first, fifth, and tenth explants cultures were detected.
The results showed that the culture cycle of this new method (Cell Isolation: 2.67 ± 0.86 days, Explants removal: 8.83 ± 0.79 days, Cell Passage: 15.17 ± 1.39 days) was significantly shorter than that of the traditional method (Cell Isolation: 8.67 ± 1.84 days, Explants removal: 17.67 ± 2.17 days, Cell Passage: 22.67 ± 1.84 days). No significant difference was observed between the phenotypes of the fibroblasts obtained from the first explants culture and cultures less than 10 times (p > 0.05).
Taken together, this study provides an effective method for the primary culture of KFs with a higher specimen utilization rate and shorter culture cycle.
This method breaks through the limitation of traditional explants culture requiring a large number of keloid specimens and provides a rich source of KFs for the study of keloid.
瘢痕疙瘩（KD）是一种复杂的纤维增生性疾病，但其发病机制的确切原因仍有待阐明。体外培养原发性瘢痕疙瘩成纤维细胞（KFs）一直是研究瘢痕疙瘩发病机制的一项基本措施。然而，传统的原代培养方法存在一些局限性，如培养周期长、标本利用率低等。
改进瘢痕疙瘩外植体培养方法。
我们提出了一种改进的新“外植体多次培养方法”——将瘢痕疙瘩外植体重复用于原代培养，并在特定培养时间收获原代KFs。同时，检测从第一次、第五次和第十次外植体培养中获得的KFs的纯度、增殖、凋亡、迁移、侵袭、细胞外基质合成以及一些纤维化和炎症相关蛋白。
结果显示，这种新方法的培养周期（细胞分离：2.67±0.86天，外植体去除：8.83±0.79天，细胞传代：15.17±1.39天）明显短于传统方法（细胞分离：8.67±1.84天，外植体去除：17.67±2.17天，细胞传代：22.67±1.84天）。从第一次外植体培养获得的成纤维细胞与少于10次培养的成纤维细胞表型之间未观察到显著差异（p>0.05）。
综上所述，本研究为KFs的原代培养提供了一种有效的方法，具有更高的标本利用率和更短的培养周期。
该方法突破了传统外植体培养需要大量瘢痕疙瘩标本的限制，为瘢痕疙瘩的研究提供了丰富的KFs来源。
