German Center for Neurodegenerative Diseases (DZNE) Dresden, 01307, Dresden, Germany.
CRTD - Center for Regenerative Therapies Dresden, Genomics of Regeneration, Technische Universität Dresden, 01307, Dresden, Germany.
Sci Rep. 2018 May 30;8(1):8433. doi: 10.1038/s41598-018-26417-1.
Distinct micro-environmental properties have been reported to be essential for maintenance of neural precursor cells (NPCs) within the adult brain. Due to high complexity and technical limitations, the natural niche can barely be studied systematically in vivo. By reconstituting selected environmental properties (adhesiveness, proteolytic degradability, and elasticity) in geldrop cultures, we show that NPCs can be maintained stably at high density over an extended period of time (up to 8 days). In both conventional systems, neurospheres and monolayer cultures, they would expand and (in the case of neurospheres) differentiate rapidly. Further, we report a critical dualism between matrix adhesiveness and degradability. Only if both features are functional NPCs stay proliferative. Lastly, Rho-associated protein kinase was identified as part of a pivotal intracellular signaling cascade controlling cell morphology in response to environmental cues inside geldrop cultures. Our findings demonstrate that simple manipulations of the microenvironment in vitro result in an important preservation of stemness features in the cultured precursor cells.
已有研究报道，在成人大脑内维持神经前体细胞（NPCs）需要独特的微环境特性。由于其复杂性和技术限制，天然小生境在体内几乎无法进行系统研究。通过在微滴培养中重建特定的环境特性（粘附性、蛋白水解降解性和弹性），我们发现 NPCs 可以在高细胞密度下稳定地维持较长时间（长达 8 天）。在传统的培养系统中，神经球和单层培养中，它们会迅速扩增和（在神经球的情况下）分化。此外，我们报告了基质粘附性和可降解性之间的关键二元性。只有当这两个特性都起作用时，NPCs 才会保持增殖状态。最后，Rho 相关蛋白激酶被鉴定为细胞内信号转导级联反应的一部分，该级联反应控制细胞形态对微滴培养中环境信号的反应。我们的研究结果表明，体外对微环境的简单操作可使培养的前体细胞重要的干性特征得到重要的保留。
