Terek Julia C, Hebb Matthew O, Flynn Lauren E
School of Biomedical Engineering, The University of Western Ontario, London, OntarioN6A 5B9, Canada.
Department of Clinical Neurological Sciences, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, OntarioN6A 5A5, Canada.
ACS Pharmacol Transl Sci. 2023 Jan 17;6(2):320-333. doi: 10.1021/acsptsci.2c00232. eCollection 2023 Feb 10.
Biomaterials derived from brain extracellular matrix (ECM) have the potential to promote neural tissue regeneration by providing instructive cues that can direct cell survival, proliferation, and differentiation. This study focused on the development and characterization of microcarriers derived from decellularized brain tissue (DBT) as a platform for neural progenitor cell culture. First, a novel detergent-free decellularization protocol was established that effectively reduced the cellular content of porcine and rat brains, with a >97% decrease in the dsDNA content, while preserving collagens (COLs) and glycosaminoglycans (GAGs). Next, electrospraying methods were applied to generate ECM-derived microcarriers incorporating the porcine DBT that were stable without chemical cross-linking, along with control microcarriers fabricated from commercially sourced bovine tendon COL. The DBT microcarriers were structurally and biomechanically similar to the COL microcarriers, but compositionally distinct, containing a broader range of COL types and higher sulfated GAG content. Finally, we compared the growth, phenotype, and neurotrophic factor gene expression levels of rat brain-derived progenitor cells (BDPCs) cultured on the DBT or COL microcarriers within spinner flask bioreactors over 2 weeks. Both microcarrier types supported BDPC attachment and expansion, with immunofluorescence staining results suggesting that the culture conditions promoted BDPC differentiation toward the oligodendrocyte lineage, which may be favorable for cell therapies targeting remyelination. Overall, our findings support the further investigation of the ECM-derived microcarriers as a platform for neural cell derivation for applications in regenerative medicine.
源自脑细胞外基质(ECM)的生物材料有潜力通过提供可指导细胞存活、增殖和分化的诱导信号来促进神经组织再生。本研究聚焦于源自脱细胞脑组织(DBT)的微载体的开发与特性表征,将其作为神经祖细胞培养的平台。首先,建立了一种新型的无洗涤剂脱细胞方案,该方案有效降低了猪脑和大鼠脑的细胞含量,双链DNA含量减少>97%,同时保留了胶原蛋白(COLs)和糖胺聚糖(GAGs)。接下来,应用电喷雾法生成包含猪DBT的源自ECM的微载体,这些微载体无需化学交联即可稳定存在,同时还制备了由市售牛肌腱COL制成的对照微载体。DBT微载体在结构和生物力学上与COL微载体相似,但组成不同,含有更广泛的COL类型和更高的硫酸化GAG含量。最后,我们比较了在旋转瓶生物反应器中培养2周的大鼠脑源性祖细胞(BDPCs)在DBT或COL微载体上的生长、表型和神经营养因子基因表达水平。两种微载体类型均支持BDPC的附着和扩增,免疫荧光染色结果表明培养条件促进BDPC向少突胶质细胞谱系分化,这可能有利于针对髓鞘再生的细胞治疗。总体而言,我们的研究结果支持进一步研究源自ECM的微载体作为神经细胞衍生平台在再生医学中的应用。