Department of Animal Physiology and Endocrinology, University of Agriculture in Krakow, Krakow, Poland.
Department of Animal Reproduction, Anatomy and Genomics, University of Agriculture in Krakow, Krakow, Poland.
Biotechnol Prog. 2021 Jul;37(4):e3152. doi: 10.1002/btpr.3152. Epub 2021 Apr 9.
Cultured fibroblast cells, especially dermal cells, are used for various types of scientific research, particularly within the medical field. Desirable features of the cells include their ease of isolation, rapid cellular growth, and high degree of robustness. Currently, fibroblasts are mainly used to obtain pluripotent cells via a reprogramming process. Dermal fibroblasts, are particularly useful for gene therapies used for promoting wound healing or minimizing skin aging. In recent years, fibroblast transfection efficiencies have significantly improved. In order to introduce molecules (most often DNA or RNA) into cells, viral-based systems (transduction) or non-viral methods (transfection) that include physical/mechanical processes or lipid reagents may be used. In this article, we describe critical points that should be considered when selecting a method for transfecting fibroblasts. The most effective methods used for the transfection of fibroblasts include both viral-based and non-viral nucleofection systems. These methods result in a high level of transgene expression and are superior in terms of transfection efficacy and viability.
培养的成纤维细胞,尤其是皮肤成纤维细胞,被广泛应用于各种类型的科学研究,尤其是在医学领域。这些细胞的理想特性包括易于分离、快速细胞生长和高度稳健性。目前,成纤维细胞主要用于通过重编程过程获得多能细胞。皮肤成纤维细胞特别适用于用于促进伤口愈合或最小化皮肤衰老的基因治疗。近年来,成纤维细胞的转染效率有了显著提高。为了将分子(通常是 DNA 或 RNA)导入细胞,可使用基于病毒的系统(转导)或非病毒方法(转染),包括物理/机械过程或脂质试剂。在本文中,我们描述了在选择转染成纤维细胞的方法时应考虑的关键点。转染成纤维细胞最有效的方法包括基于病毒的和非病毒的核转染系统。这些方法可实现高水平的转基因表达,在转染效率和细胞活力方面具有优势。