Department of Internal Medicine and Infectious Diseases-Coronary Care Unit, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy; Laboratory of Experimental Cardiology for Cell and Molecular Therapy, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy; Department of Molecular Medicine, Unit of Cardiology, University of Pavia, Italy.
Department of Internal Medicine and Infectious Diseases-Coronary Care Unit, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy; Laboratory of Experimental Cardiology for Cell and Molecular Therapy, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy.
Pharmacol Res. 2018 Jan;127:49-57. doi: 10.1016/j.phrs.2017.11.014. Epub 2017 Nov 14.
Mesenchymal stromal cells are excellent candidates for regenerative medicine since they are multipotent, easy to isolate, can be expanded to obtain clinically relevant numbers and are immunoprivileged. Stable genetic modification with viral vectors can improve mesenchymal stromal cell function and enhance their therapeutic potential. However, standard viral vectors achieve sub-optimal transduction efficiency with a single infection. On the other hand, multiple transduction cycles or antibiotic-based selection methods may alter the stem cell phenotype. We hypothesized that the use of lentiviral vectors containing specific regulatory sequences may result in improved transduction efficiency. Thus, we compared two types of third generation lentiviral vectors, one of which, the pLenti7.3 vector, contains the optimized sequences for Polypurine Tract and Woodchuck Post-transcriptional Regulatory Element. We demonstrated that with the pLenti7.3 it is possible to efficiently transduce human mesenchymal stromal cells with a single transduction cycle. Additionally, we successfully showed that by using the pLenti7.3 vector it is possible to efficiently over-express different growth factors, particularly relevant for cardiac protection and differentiation, in human mesenchymal stromal cells.
间充质基质细胞是再生医学的优秀候选者，因为它们具有多能性、易于分离、可以扩增以获得临床相关数量，并且具有免疫特权。使用病毒载体进行稳定的基因修饰可以改善间充质基质细胞的功能并增强其治疗潜力。然而，标准的病毒载体在单次感染时只能达到亚最佳的转导效率。另一方面，多次转导周期或基于抗生素的选择方法可能会改变干细胞表型。我们假设使用含有特定调节序列的慢病毒载体可能会导致转导效率的提高。因此，我们比较了两种类型的第三代慢病毒载体，其中一种是 pLenti7.3 载体，它包含优化的 Polypurine Tract 和 Woodchuck Post-transcriptional Regulatory Element 序列。我们证明，使用 pLenti7.3 载体可以在单次转导周期内有效地转导人骨髓基质细胞。此外，我们还成功地表明，使用 pLenti7.3 载体可以有效地在人骨髓基质细胞中过表达不同的生长因子，特别是与心脏保护和分化相关的生长因子。
