Department of Orthopedic Surgery, West China Hospital, Sichuan University; Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, West China Hospital, Sichuan University, Chengdu, 610041, China.
Division of Health Quarantine, Shenzhen Entry-Exit Inspection and Quarantine Bureau, 518045, China.
J Biomed Mater Res A. 2018 Mar;106(3):686-697. doi: 10.1002/jbm.a.36264. Epub 2017 Nov 6.
Stem cell-based gene therapy has been considered in the treatment of many degenerative diseases. Gene-modified stem cells should maintain its reproductive activity without losing stem cell properties, including genetic phenotype and differentiation potential. In the study, a novel poly (amidoamine) with pendant aminobutyl group (PAA-BA) designed by our group was used in the transfection of bone marrow mesenchymal stromal cells (BMSCs) and the cellular properties post-transfection were evaluated, including DNA content, colony forming capacity, genetic phenotype, and multi-directional differentiation. Two classical non-viral gene delivery vectors, polyethylenimine (PEI) and Lipofectamine 2000 (LP2000) were also used. Compared to non-transfected group, PAA-BA showed minor decreased DNA content but maintained BMSCs' phenotype, reproductive activity and multi-differentiation potential (osteogenic, chondrogenic, adipogenic, and neurogenic differentiation). Both PAA-BA and PEI transfected BMSCs demonstrated improved osteogenic differentiation ability at late stage but suppressed adipogenic as well as mature neural differentiation in vitro. LP2000 and PEI transfected BMSCs displayed significantly lower DNA content and reproductive activity. These findings suggest that PAA-BA is one of safe gene delivery vectors in BMSCs transfection and plays a role in stem cell's osteogenic and neurogenic differentiation. This study proposes the potential application of PAA-BA in BMSCs based gene therapy, in particular bone and nerve relative diseases. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 686-697, 2018.
基于干细胞的基因治疗已被应用于许多退行性疾病的治疗中。基因修饰后的干细胞应在保持其生殖活性的同时，不丧失干细胞特性，包括遗传表型和分化潜能。在本研究中，我们设计了一种新型的带有支链的聚（酰胺-胺）（PAA-BA），用于转染骨髓间充质基质细胞（BMSCs），并对转染后细胞的特性进行了评估，包括 DNA 含量、集落形成能力、遗传表型和多向分化。同时还使用了两种经典的非病毒基因传递载体：聚乙烯亚胺（PEI）和脂质体 2000（LP2000）。与未转染组相比，PAA-BA 转染后细胞的 DNA 含量略有降低，但保持了 BMSCs 的表型、生殖活性和多向分化潜能（成骨、成软骨、成脂和成神经分化）。PAA-BA 和 PEI 转染的 BMSCs 在晚期均表现出增强的成骨分化能力，但在体外抑制了成脂分化和成熟的神经分化。LP2000 和 PEI 转染的 BMSCs 的 DNA 含量和生殖活性显著降低。这些结果表明，PAA-BA 是一种安全的 BMSCs 转染基因传递载体，可促进干细胞的成骨和神经分化。本研究提出了 PAA-BA 在基于 BMSCs 的基因治疗中的潜在应用，特别是在骨和神经相关疾病的治疗中。 © 2017 Wiley Periodicals, Inc. J 生物材料研究杂志 A 部分：106A：686-697，2018 年。
