期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 159, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/61037
关键词
Bioengineering; Issue 159; hypertrophic scar; fibroblasts; macromolecular crowding; collagen; extracellular matrix; density gradient medium; polyvinylpyrrolidone
资金
- Singapore's Agency for Science, Technology and Research [SPF 2013/004, IAF-PP/2017, H17/01/a0/009]
It has been shown that in vivo tissues are highly crowded by proteins, nucleic acids, ribonucleoproteins, polysaccharides, etc. The following protocol applies a macromolecular crowding (MMC) technique to mimic this physiological crowding through the addition of neutral polymers (crowders) to cell cultures in vitro. Previous studies using Ficoll or dextran as crowders demonstrate that the expression of collagen I and fibronectin in WI38 and WS-1 cell lines are significantly enhanced using the MMC technique. However, this technique has not been validated in primary hypertrophic scar-derived human skin fibroblasts (hHSFs). As hypertrophic scarring arises from the excessive deposition of collagen, this protocol aims to construct a collagen-rich in vitro hypertrophic scar model by applying the MMC technique with hHSFs. This optimized MMC model has been shown to possess more similarities with in vivo scar tissue compared to traditional 2-dimensional (2-D) cell culture systems. In addition, it is cost-effective, time-efficient, and ethically desirable compared to animal models. Therefore, the optimized model reported here offers an advanced in vivo-like model for hypertrophic scar-related studies.
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