An Efficient Method for the Isolation and Cultivation of Hypothalamic Neural Stem/Progenitor Cells From Mouse Embryos.

Yichao Ou,Peirong Niu,Xingqin Wang,Mingfeng Zhou,Mengjie Che,Kai Li,Guangsen Wu,Zhanpeng Feng,Songtao Qi,Junjie Peng,Haodong Gong

doi:10.3389/fnana.2022.711138

Yichao Ou, Peirong Niu + Show 9 more

Open Access

https://doi.org/10.3389/fnana.2022.711138

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Abstract

The hypothalamus is the key region that regulates the neuroendocrine system as well as instinct behaviors, and hypothalamic dysfunction causes refractory clinical problems. Recent studies have indicated that neural stem/progenitor cell (NSPC) in the hypothalamus play a crucial role in hypothalamic function. However, specific hypothalamic NSPC culture methods have not been established, especially not detailed or efficient surgical procedures. The present study presented a convenient, detailed and efficient method for the isolation and cultivation of hypothalamic NSPCs from embryonic day 12.5 mice. The procedure includes embryo acquisition, brain microdissection to quickly obtain hypothalamic tissue and hypothalamic NSPC culture. Hypothalamic NSPCs can be quickly harvested and grow well in both neurosphere and adherent cultures through this method. Additionally, we confirmed the cell origin and evaluated the proliferation and differentiation properties of cultured cells. In conclusion, we present a convenient and practical method for the isolation and cultivation of hypothalamic NSPCs that can be used in extensive hypothalamic studies.

Highlights

The hypothalamus is one of the most important and complicated parts of the brain, and it regulates various essential biological processes, including energy homeostasis, maintenance of the internal environment, survival behavior, thermoregulation, aging, and circadian rhythms (Sternson, 2013; Saper and Lowell, 2014)
To understand the development of hypothalamus and the role of neurogenesis in hypothalamic function regulation, it is critical to culture and study hypothalamic neural stem/progenitor cell (NSPC)
C57BL/6 female mice that were pregnant with 12.5-day-old embryos were purchased from the Experimental Animal Center of Southern Medical University (Guangzhou, China) to generate hypothalamic NSPC cultures

Summary

Introduction

The hypothalamus is one of the most important and complicated parts of the brain, and it regulates various essential biological processes, including energy homeostasis, maintenance of the internal environment, survival behavior, thermoregulation, aging, and circadian rhythms (Sternson, 2013; Saper and Lowell, 2014). Hypothalamic injuries may lead to refractory complications, especially central diabetes insipidus, obesity, growth retardation in children, and gonadal dysfunction (Honegger et al, 1999; Feng et al, 2018). In addition to exogenous supplementation for certain hormone deficiencies, an efficient method to improve hypothalamic dysfunction is still unclear. A few studies have indicated that complicated hypothalamic function can be precisely modulated by neurogenesis in the hypothalamus (Czupryn et al, 2011; Zhang et al, 2017). Efforts are needed to determine how hypothalamic neural stem/progenitor cell (NSPCs) regulate hypothalamic function and how NSPCs communicate with surrounding cells in the Method for Hypothalamic NSPC Culture hypothalamus. To understand the development of hypothalamus and the role of neurogenesis in hypothalamic function regulation, it is critical to culture and study hypothalamic NSPCs

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Results

Conclusion