de Carvalho Talita Giacomet, Pellenz Felipe Matheus, Laureano Alvaro, da Rocha Silla Lucia Mariano, Giugliani Roberto, Baldo Guilherme, Matte Ursula
Gene Therapy Center, Experimental Research Center, Hospital de Clínicas de Porto Alegre, Rua Ramiro Barcelos, 2350, Porto Alegre, RS, 90035-903, Brazil.
Post-Graduation Program on Genetics and Molecular Biology, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
Biotechnol Lett. 2018 Mar;40(3):617-622. doi: 10.1007/s10529-018-2505-8. Epub 2018 Jan 17.
Mesenchymal stromal cells (MSCs) are potential targets for cell and gene therapy-based approaches against a variety of different diseases. The MSCs from bone marrow are a promising target population as they are capable of differentiating along multiple lineages and have significant expansion capability. These characteristics make them strong candidates for delivering genes and restoring organ systems function. However, as other primary cells, MSCs are difficult to transfect. In order to standardize a simple protocol for transfection of MSCs, we conducted a series of experiments and achieved a protocol that does not require the use of viral particles or specific expensive equipment.
MSCs transfection at early passages using a ratio lipid/DNA of 3.0 µL/µg with Lipofectamine 3000 yields good transfection efficiencies for human MSCs (up to 26%) and is rapid, simple, and safe.
间充质基质细胞（MSCs）是基于细胞和基因治疗方法对抗多种不同疾病的潜在靶点。来自骨髓的MSCs是一个有前景的目标群体，因为它们能够沿多个谱系分化且具有显著的扩增能力。这些特性使其成为递送基因和恢复器官系统功能的有力候选者。然而，与其他原代细胞一样，MSCs难以转染。为了标准化一种简单的MSCs转染方案，我们进行了一系列实验，并获得了一种无需使用病毒颗粒或特定昂贵设备的方案。
使用Lipofectamine 3000以脂质/DNA比例为3.0 μL/μg在早期传代时对MSCs进行转染，可为人MSCs产生良好的转染效率（高达26%），且快速、简单、安全。
