Animal Physiology and Reproduction Division, ICAR-Central Institute for Research on Goats, Makhdoom, Farah, Mathura, Uttar Pradesh, 281 122, India.
Department of Biotechnology, GLA University, Mathura, Uttar Pradesh, 281 406, India.
Sci Rep. 2023 Aug 22;13(1):13716. doi: 10.1038/s41598-023-38634-4.
The enhanced availability of functional fibroblasts from precious tissue samples requires an ideal cell-culture system. Therefore, this study was designed to investigate the performance of caprine adult fibroblast cells (cadFibroblast) when cultivated in different culture media. The cadFibroblast cell lines from adult Barbari (Capra hircus) bucks were established and the effect of different media viz. DMEM/F-12 [with low-glucose (5.5 mM; DL) and high-glucose (30 mM; DH)], α-MEM [with low-glucose (5.5 mM; ML) and with high-glucose (30 mM; MH)], and fibroblast growth medium (FGM) were evaluated. Cells were then compared for growth characteristics and in-vitro dynamics through cellular morphology, proliferation, population-doubling time, double-immunocytochemistry, colony-forming units, wound healing, transwell migration, and differential expression of fibroblast-specific markers (FSP-1 and vimentin). The results of immunocytochemistry, transwell migration/invasion, and wound healing assays showed the superiority of DH over DL and other media tested. Whereas, similar effects of glucose supplementation and expression of FSP-1 were not observed in α-MEM. Transwell migration was significantly (p < 0.05) lower in FGM compared with other media tested. Overall, our results illustrate the media-dependent deviation in in-vitro dynamics and culture characteristics of cadFibroblasts that may be useful to develop strategies to cultivate these cells efficiently for research and downstream applications.
从珍贵的组织样本中获得功能齐全的成纤维细胞需要一个理想的细胞培养系统。因此,本研究旨在研究山羊成纤维细胞(cadFibroblast)在不同培养介质中培养时的性能。本研究从成年公山羊(Capra hircus)中建立了 cadFibroblast 细胞系,并研究了不同培养基(DMEM/F-12 [含低葡萄糖(5.5mM;DL)和高葡萄糖(30mM;DH)]、α-MEM [含低葡萄糖(5.5mM;ML)和高葡萄糖(30mM;MH)]和成纤维细胞生长培养基(FGM))对细胞生长特性和体外动力学的影响。通过细胞形态、增殖、倍增时间、双免疫细胞化学、集落形成单位、伤口愈合、transwell 迁移和成纤维细胞特异性标志物(FSP-1 和波形蛋白)的差异表达,比较了细胞。免疫细胞化学、transwell 迁移/侵袭和伤口愈合试验的结果表明,DH 优于 DL 和其他测试的培养基。然而,在α-MEM 中没有观察到葡萄糖补充和 FSP-1 表达的类似效果。与其他测试的培养基相比,FGM 中的 transwell 迁移明显(p < 0.05)较低。总体而言,我们的结果说明了 cadFibroblast 体外动力学和培养特性的培养基依赖性偏差,这可能有助于制定策略来有效地培养这些细胞,用于研究和下游应用。