Budiariati Vista, Rinendyaputri Ratih, Noviantari Ariyani, Budiono Dwi, Fahrudin Mokhamad, Juliandi Berry, Boediono Arief
Department of Anatomy, Physiology, and Pharmacology, Faculty of Veterinary Medicine, IPB University, Bogor, Indonesia.
Center for Research and Development of Biomedical and Basic Health Technology, National Institute of Health Research and Development, Ministry of Health Republic of Indonesia.
J Stem Cells Regen Med. 2019 Dec 24;15(2):35-44. doi: 10.46582/jsrm.1502008. eCollection 2019.
Conditioned medium has now gained increasing interest since the development of secretome-based therapy. Various types of cells have been studied as a source of the secretome. One of them is neural progenitor cells (NPCs). These are cells that capable of differentiating into neurons as well as glial cells. Indeed, the study on NPCs has risen in the last few decades, but the study on the differentiated cells has not clearly described. The most common procedures that widely used to get the conditioned medium is starvation. However, cell starvation may cause environmental stress and become an apoptotic trigger for the cells. In this study, we analyzed the effect of starvation on differentiated cells from E17 rat neural progenitor cells (NPCs) based on cells characteristics and secretome profile. We found that starvation decreased cells viability and affected the heterogeneity of the cell population. Astrocytes survived more under nutrient deprivation conditions, and the progenitor cells showed a higher tendency to differentiate to glial cells than neurons. Duration of starvation also influenced the secretome profile, alterations found in protein types and also their function in the biological process. During 24 hours of starvation, cells secreted proteins that were used to maintain cell growth, stimulate differentiation, and produce energy, but there were also proteins that identified and involved in autophagy activation. After 48 hours of starvation, astrocytes that became the dominant cells secreted proteins that try to keep protecting the remaining neurons.
自基于分泌组的疗法发展以来,条件培养基现在越来越受到关注。各种类型的细胞已被研究作为分泌组的来源。其中之一是神经祖细胞(NPCs)。这些细胞能够分化为神经元以及神经胶质细胞。事实上,在过去几十年中对NPCs的研究有所增加,但对分化细胞的研究尚未得到清晰描述。广泛用于获取条件培养基的最常见方法是饥饿处理。然而,细胞饥饿可能会导致环境应激并成为细胞凋亡的触发因素。在本研究中,我们基于细胞特征和分泌组谱分析了饥饿对E17大鼠神经祖细胞(NPCs)分化细胞的影响。我们发现饥饿降低了细胞活力并影响了细胞群体的异质性。在营养剥夺条件下,星形胶质细胞存活得更多,并且祖细胞向神经胶质细胞分化的趋势高于向神经元分化的趋势。饥饿持续时间也影响分泌组谱,在蛋白质类型及其在生物过程中的功能方面都发现了变化。在饥饿24小时期间,细胞分泌用于维持细胞生长、刺激分化和产生能量的蛋白质,但也有被鉴定并参与自噬激活的蛋白质。在饥饿48小时后,成为优势细胞的星形胶质细胞分泌试图继续保护剩余神经元的蛋白质。