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胶质细胞条件培养基诱导人脂肪间充质干细胞的神经分化。

Neural differentiation of human adipose-derived mesenchymal stem cells induced by glial cell conditioned media.

机构信息

Department of Biomedical and Biotechnological Sciences, Section of Physiology, University of Catania, Catania, Italy.

Department of Clinical and Experimental Medicine, University of Catania, Catania, Italy.

出版信息

J Cell Physiol. 2018 Oct;233(10):7091-7100. doi: 10.1002/jcp.26632. Epub 2018 May 8.

Abstract

Adipose-derived mesenchymal stem cells (ASCs) may transdifferentiate into cells belonging to mesodermal, endodermal, and ectodermal lineages. The aim of this study was to verify whether a neural differentiation of ASCs could be induced by a conditioned medium (CM) obtained from cultures of olfactory ensheathing cells (OECs) or Schwann cells (SCs). ASCs were isolated from the stromal vascular fraction of adipose tissue and expanded for 2-3 passages. They were then cultured in OEC-CM or SC-CM for 24 hr or 7 days. At each stage, the cells were tested by immunocytochemistry and flow cytometer analysis to evaluate the expression of typical neural markers such as Nestin, PGP 9.5, MAP2, Synapsin I, and GFAP. Results show that both conditioned media induced similar positive effects, as all tested markers were overexpressed, especially at day 7. Overall, an evident trend toward neuronal or glial differentiation was not clearly detectable in many cases. Nevertheless, a higher tendency toward a neuronal phenotype was recognized for OEC-CM (considering MAP2 increases). On the other hand, SC-CM would be responsible for a more marked glial induction (considering GFAP increases). These findings confirm that environmental features can induce ASCs toward a neural differentiation, either as neuronal or glial elements. Rather than supplementing the culture medium by adding chemical agents, a "more physiological" condition was obtained here by means of soluble factors (cytokines/growth factors) likely released by glial cells. This culture strategy may provide valuable information in the development of cell-based therapeutic approaches for pathologies affecting the central/peripheral nervous system.

摘要

脂肪间充质干细胞(ASCs)可转分化为中胚层、内胚层和外胚层谱系的细胞。本研究旨在验证 ASC 是否可通过嗅鞘细胞(OECs)或雪旺细胞(SCs)培养获得的条件培养基(CM)诱导神经分化。从脂肪组织基质血管部分分离 ASC 并扩增 2-3 代。然后将其在 OEC-CM 或 SC-CM 中培养 24 小时或 7 天。在每个阶段,通过免疫细胞化学和流式细胞仪分析测试细胞,以评估神经标志物如巢蛋白、PGP 9.5、MAP2、突触素 I 和 GFAP 的表达。结果表明,两种条件培养基均诱导出相似的积极效果,所有测试的标志物均过度表达,尤其是在第 7 天。总体而言,许多情况下并未明显检测到向神经元或神经胶质分化的明显趋势。然而,OEC-CM 更倾向于诱导神经元表型(考虑到 MAP2 增加)。另一方面,SC-CM 将负责更明显的神经胶质诱导(考虑到 GFAP 增加)。这些发现证实环境特征可诱导 ASC 向神经分化,无论是神经元还是神经胶质细胞。本研究通过使用可能由神经胶质细胞释放的可溶性因子(细胞因子/生长因子)替代在培养基中添加化学试剂的方法,获得了一种“更生理”的条件,从而为影响中枢/周围神经系统的病理的细胞治疗方法的发展提供了有价值的信息。

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