瘢痕疙瘩成纤维细胞低密度培养的克隆能力分析

Clonal Analysis on Low Density Culture of Keloid Fibroblasts

目的比较瘢痕疙瘩成纤维细胞(KFs)和正常真皮成纤维细胞(NFs)间克隆形成能力的差异,探讨瘢痕疙瘩中病理性干细胞是否存在,及其对瘢痕疙瘩发生发展的影响。方法利用酶消化法获得瘢痕疙瘩和正常皮肤组织的原代细胞,以4 000个/皿的密度接种,进行低密度培养,2周后观察细胞克隆的形成及形态变化。结果低密度培养条件下的瘢痕疙瘩成纤维细胞和正常皮肤成纤维细胞都可形成克隆,但KFs可形成明显的克隆集落,NFs形成的克隆不明显且松散。KFs克隆形成率高于NFs,为(0.80±0.21)%,而NFs为(0.18±0.06)%,两者间差异显著(P<0.05)。结论低密度培养条件下,瘢痕疙瘩成纤维细胞的克隆形成能力高于正常皮肤成纤维细胞,可能与瘢痕疙瘩组织中存在病理性瘢痕疙瘩干细胞有关。

Objective To compare the clone forming capacity between keloid fibroblasts （KFs） and normal dermal fibroblasts （NFs）, and to explore whether there are pathological stem cells in keloid and the role of keloid stem cells on the mechanism of keloid formation. Methods Primary cells were obtained from keloid tissue and normal dermis respectively through enzyme digestion method, and then were cultured under low-density conditions of 4 000/dish. Two weeks later, the clone formation and morphology were determined. Results Under low-density culture conditions, clones were formed in both KFs and NFs. KFs formed distinct and dense clones while NFs formed loose and fewer ones. KFs exhibited higher clone forming efficiency （CFE） of （0.80±0.21）%than NFs with CFE of （0.18±0.06）% （P〈0.05）. Conclusion With low-density culture, keloid fibroblasts possess higher level of cloning capacity than normal dermal fibroblasts, which suggests pathological stem cells may exist in keloid tissues.