Conti Luciano, Cattaneo Elena
Department of Pharmacological Sciences and Center for Stem Cell Research, University of Milan, Milan, Italy.
Methods Mol Biol. 2008;438:319-32. doi: 10.1007/978-1-59745-133-8_24.
Transplantation of neural cells engineered to produce growth factors or molecules with antitumor effects have the potential of grafted cells to be used as vectors for protein delivery in animal models of diseases. In this context, neural stem cells (NSCs), since their identification, have been considered an attractive subject for therapeutic applications to the damaged brain. NSCs have been shown to include attributes important for potential successful ex vivo gene therapy approaches: they show extensive in vitro expansion and, in some cases, a particular tropism toward pathological brain areas. Clearly, the challenges for future clinical development of this approach are in the definition of the most appropriate stem cells for a given application, what genes or chemicals can be delivered, and what diseases are suitable targets. Ideally, NSC lines should be homogeneous and well characterized in terms of their in vitro stability and grafting capacity. We discuss two possible approaches to produce homogeneous and stable progenitor and NSC lines that exploit an oncogene-based immortalization, or, in the second case, a novel protocol for growth factor expansion of stem cells with radial glia-like features. Furthermore, we describe the use of retroviral particles for genetic engineering.
经基因工程改造以产生生长因子或具有抗肿瘤作用分子的神经细胞移植，其移植细胞有潜力在疾病动物模型中用作蛋白质递送载体。在此背景下，神经干细胞（NSCs）自被发现以来，就被认为是受损大脑治疗应用的一个有吸引力的研究对象。神经干细胞已被证明具有对潜在成功的体外基因治疗方法很重要的特性：它们在体外能大量扩增，并且在某些情况下，对病理性脑区有特定的趋向性。显然，这种方法未来临床开发面临的挑战在于确定针对特定应用最合适的干细胞、可递送哪些基因或化学物质以及哪些疾病是合适的靶点。理想情况下，神经干细胞系应在体外稳定性和移植能力方面具有同质性且特征明确。我们讨论了两种产生同质性和稳定祖细胞及神经干细胞系的可能方法，一种利用基于癌基因的永生化，另一种则是一种用于具有放射状胶质细胞样特征的干细胞生长因子扩增的新方案。此外，我们描述了逆转录病毒颗粒在基因工程中的应用。
