Aquatic Animal Health Laboratory (OIE Reference for WTD), PG & Research Department of Zoology, C. Abdul Hakeem College, Melvisharam, Vellore District, India.
J Fish Dis. 2019 Apr;42(4):573-584. doi: 10.1111/jfd.12965. Epub 2019 Feb 14.
The goal of this study was to develop and characterize a cell line from the caudal fin tissue of zebrafish and also its application as an in vitro model to study the effect of H O in wound healing. Fibroblastic cell line was developed using explant culture method from caudal fin tissue of zebrafish and characterized. This cell line was named as DrF cell line. The DrF cells treated with 0-10 µM/ml H O were tested for viability, proliferation and motility by MTT assay, trypan blue assay and chemotaxis assay, respectively. Among the different concentrations of H O , 4 µM was found to be nontoxic to study cell migration in in vitro scratch wound assay. Furthermore, the expression of proliferating cell nuclear antigen (PCNA) and chemokine receptor (CXCR4) genes was carried by qPCR. The cell survival, proliferation and migration were extremely enriched at 4 µM level of H O . We observed accelerated wound closure in DrF cells treated with H O The qPCR results indicated that H O markedly up-regulated mRNA expression of PCNA and CXCR4. The findings from our study suggest that H O at low levels promotes cell survival, proliferation, migration and wound healing in DrF cells.
本研究的目的是从斑马鱼的尾鳍组织中开发并鉴定一种细胞系,并将其作为体外模型应用于研究 H2O2对伤口愈合的影响。采用组织块培养法从斑马鱼尾鳍组织中分离出成纤维细胞系,并对其进行了鉴定。该细胞系命名为 DrF 细胞系。通过 MTT 检测、台盼蓝检测和趋化性检测,分别检测 0-10µM/ml H2O2处理的 DrF 细胞的活力、增殖和迁移能力。在不同浓度的 H2O2中,发现 4µM 对研究体外划痕伤口愈合试验中的细胞迁移无毒性。此外,通过 qPCR 检测增殖细胞核抗原(PCNA)和趋化因子受体(CXCR4)基因的表达。在 4µM 水平的 H2O2下,细胞的存活、增殖和迁移得到了极大的富集。我们观察到 DrF 细胞在 H2O2处理后伤口愈合加快。qPCR 结果表明,H2O2显著上调了 PCNA 和 CXCR4 的 mRNA 表达。本研究结果表明,低水平的 H2O2促进 DrF 细胞的存活、增殖、迁移和伤口愈合。