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间充质干细胞条件培养基对角膜基质成纤维细胞创面愈合活性的影响。

The effect of mesenchymal stem cell conditioned media on corneal stromal fibroblast wound healing activities.

机构信息

Department of Pathology, School of Medical Sciences, University of New South Wales, Randwick, New South Wales, Australia.

出版信息

Br J Ophthalmol. 2010 Aug;94(8):1067-73. doi: 10.1136/bjo.2009.165837. Epub 2009 Dec 3.

Abstract

AIMS

To investigate the effects of conditioned media from mesenchymal stem cells (MSC) on the wound healing activities of corneal stromal fibroblasts.

METHODS

Cell cycle analysis and early stage activation of apoptosis, chemotactic chambers and fibroblast-populated type I collagen gels were used to assess corneal stromal fibroblast proliferation, migration and contraction, respectively. Fibroblasts were obtained from human donor corneas and MSC from fresh rat bone marrow. MSC conditioned media and fibroblast culture medium (FCM), with and without calf serum supplementation, were compared.

RESULTS

MSC conditioned media and serum-free FCM had an inhibitory effect on the progression of corneal fibroblasts through the cell cycle. There was a significant increase in the number of cells in the G0-G1 phase for MSC conditioned media and serum-free FCM (p=0.001, p=0.97 respectively). Fibroblast migration and relaxed and stressed gel contraction were significantly inhibited by MSC conditioned media and serum-free FCM compared with FCM with serum (all p=0.001). Glucose and lactate analysis confirmed that these factors were not contributing to this effect.

CONCLUSION

MSC conditioned media was found to inhibit the wound healing activities of corneal stromal fibroblasts in vitro. Putative factors secreted by MSC could be developed for therapeutic use in corneal repair.

摘要

目的

研究间充质干细胞(MSC)条件培养基对角膜基质成纤维细胞伤口愈合活性的影响。

方法

采用细胞周期分析和早期凋亡激活、趋化室和纤维母细胞填充 I 型胶原凝胶,分别评估角膜基质成纤维细胞的增殖、迁移和收缩。成纤维细胞取自人供体角膜,MSC 取自新鲜大鼠骨髓。比较 MSC 条件培养基和无小牛血清的成纤维细胞培养基(FCM),以及添加和不添加小牛血清的条件培养基和 FCM。

结果

MSC 条件培养基和无血清 FCM 通过细胞周期对角膜成纤维细胞的进展具有抑制作用。MSC 条件培养基和无血清 FCM 中 G0-G1 期细胞数量明显增加(p=0.001,p=0.97)。与含血清的 FCM 相比,MSC 条件培养基和无血清 FCM 显著抑制了纤维母细胞的迁移以及松弛和紧张凝胶的收缩(均 p=0.001)。葡萄糖和乳酸分析证实,这些因素并非导致这种抑制作用的原因。

结论

MSC 条件培养基被发现可抑制体外角膜基质成纤维细胞的伤口愈合活性。MSC 分泌的潜在因子可开发用于角膜修复的治疗用途。

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