Luo Jiao, Tang Qiaoyan, Lin Tanjing, Liu Jiabang, Wu Zhiheng, Zhang Xintao, Zhang Xiaohua, Jiang Junhai, Wang Yulong
Department of Rehabilitation Medicine, Dapeng New District Nan'ao People's Hospital, Rehabilitation Branch of the First Affiliated Hospital of Shenzhen University, Shenzhen, China.
School of Pharmacy, Guangdong Medical University, Guangdong, China.
Sci Rep. 2025 May 27;15(1):18542. doi: 10.1038/s41598-025-02847-6.
Hypothalamic neurogenesis is a complex process that plays a crucial role in neuroendocrine homeostasis, making in vivo studies of the hypothalamus particularly challenging. In this study, we present an optimized protocol for isolating and culturing hypothalamic neural stem cells (htNSCs) from neonatal (P1) mice, followed by their directed differentiation in a three-dimensional (3D) Matrigel environment. We successfully established a primary culture system that supports the stability, growth, and distinct characteristics of htNSCs. Notably, we demonstrate that htNSCs can differentiate into GnRH-like neurons within the Matrigel-based 3D culture system. These differentiated neurons exhibit typical neuronal morphology and functional characteristics. Our findings highlight the potential of neonatal htNSCs as an invaluable model for studying hypothalamic function and neurogenesis. Furthermore, this method provides a novel platform for basic research and may serve as important implications for further studying the pathological mechanism of neuroendocrine disorders in hypothalamus.
下丘脑神经发生是一个复杂的过程，在神经内分泌稳态中起关键作用，这使得下丘脑的体内研究极具挑战性。在本研究中，我们提出了一种优化方案，用于从新生（P1）小鼠中分离和培养下丘脑神经干细胞（htNSCs），随后在三维（3D）基质胶环境中进行定向分化。我们成功建立了一个支持htNSCs稳定性、生长和独特特性的原代培养系统。值得注意的是，我们证明了htNSCs可以在基于基质胶的3D培养系统中分化为促性腺激素释放激素（GnRH）样神经元。这些分化的神经元表现出典型的神经元形态和功能特征。我们的研究结果突出了新生htNSCs作为研究下丘脑功能和神经发生的宝贵模型的潜力。此外，该方法为基础研究提供了一个新平台，可能对进一步研究下丘脑神经内分泌疾病的病理机制具有重要意义。
