Bible Ellen, Chau David Y S, Alexander Morgan R, Price Jack, Shakesheff Kevin M, Modo Michel
Department of Neuroscience, Institute of Psychiatry, Kings College London, London, UK.
Nat Protoc. 2009;4(10):1440-53. doi: 10.1038/nprot.2009.156. Epub 2009 Sep 17.
Cell-replacement therapy and tissue regeneration using stem cells are of great interest to recover histological damage caused by neuro-degenerative disease or traumatic insults to the brain. To date, the main intra-cerebral delivery for these cells has been as a suspension in media through a thin needle. However, this does not provide cells with a support system that would allow tissue regeneration. Scaffold particles are needed to provide structural support to cells to form de novo tissue. In this 16-d protocol, we describe the generation and functionalization of poly (D,L-lactic-co-glycolic) acid (PLGA) particles to enhance cell attachment, the attachment procedure to avoid clumping and aggregation of cells and particles, and their preparation for intra-cerebral injection through a thin needle. Although the stem cell-scaffold transplantation is more complicated and labor-intensive than cell suspensions, it affords de novo tissue generation inside the brain and hence provides a significant step forward in traumatic brain repair.
利用干细胞进行细胞替代疗法和组织再生,对于恢复由神经退行性疾病或脑部创伤性损伤引起的组织学损伤具有重大意义。迄今为止,这些细胞主要通过细针以培养基悬浮液的形式进行脑内递送。然而,这并未为细胞提供一个能够促进组织再生的支持系统。需要支架颗粒为细胞提供结构支持以形成新生组织。在这个为期16天的实验方案中,我们描述了聚(D,L-乳酸-共-乙醇酸)(PLGA)颗粒的制备及其功能化,以增强细胞附着,避免细胞和颗粒聚集的附着程序,以及它们通过细针进行脑内注射的制备方法。尽管干细胞-支架移植比细胞悬液更复杂且劳动强度更大,但它能在脑内实现新生组织的生成,从而在创伤性脑损伤修复方面向前迈出了重要一步。