Name: Magnetic Separation and Negative Selection of Murine Neuronal Cultures (Video) | JoVE
Uploaded: 2023-08-04T13:20:00
Duration: 2 min 43 s
Description: Watch this Scientific Journal Video about Magnetic Separation and Negative Selection of Murine Neuronal Cultures at JoVE.com

magnetic separation and negative selection of murine neuronal cultures

Isolation of Targeted Hypothalamic Neurons for Studies of Hormonal, Metabolic, and Electrical Regulation

Watch this Scientific Journal Video about Magnetic Separation and Negative Selection of Murine Neuronal Cultures at JoVE.com

Magnetic Separation and Negative Selection of Murine Neuronal Cultures (Video) | JoVE 

小鼠神经元培养物的磁分离和阴性选择

首先将分离的脑细胞悬液在300G下离心八分钟。将沉淀重悬于含有0.5%BSA溶液的80微升HBSS中。加入 20 微升非神经元细胞生物素抗体混合物并孵育。用两毫升含有0.5%BSA的HBSS洗涤细胞。然后将离心机沉淀重悬于含有0.5%BSA的80微升HBSS中。现在加入20微升抗生物素微珠，并移液器充分混合。冷孵育后加入0.5毫升HBSS BSA溶液。在用0.5毫升含有0.5%BSA的HBSS冲洗色谱柱之前设置支架。接下来，在柱子下放置一个15毫升的管，并将0.5毫升的细胞悬液通过它。用0.5毫升HBSS和0.5%BSA溶液进行三次洗涤，以捕获残留的神经元细胞。从磁铁上取下色谱柱，并将其放入新的 15 毫升管中。加入一毫升含有0.5%BSA的HBSS，并使用柱塞收集磁性标记的非神经元细胞。细胞离心后，将细胞重悬于一毫升含有0.5%BSA溶液的HBSS中。在明场显微镜下对Neubauer计数室中的细胞进行计数。LepR阳性神经元在48小时后开始形成神经突。在DIV4轴突延伸显示出进展，而树突突开始出现。在DIV6，神经元充分发育。免疫荧光研究表明，没有观察到神经胶质细胞或其他非神经元细胞。细胞的神经元性质通过微管相关蛋白二染色得到证实。在DIV10时，30%的LepR阳性细胞表达POMC。在含有异质性下丘脑神经元群的一般培养物中观察到突触连接和功能。

Research

JoVE Journal

Neuroscience

Magnetic Separation and Negative Selection of Murine Neuronal Cultures

Hormonal, Metabolic, and Electrical Regulation of Isolated Hypothalamic Neurons

Hypothalamic Neural Mechanism Insights (Video) | JoVE

分离靶向下丘脑神经元，用于激素、代谢和电调节研究

The hypothalamus regulates fundamental metabolic processes by controlling functions as varied as food intake, body temperature, and hormone release. As ...

科研

神经科学

Murine Embryonal Hypothalamus Extraction, Collection, and Tissue Dissociation

小鼠胚胎下丘脑提取、采集和组织解离

Magnetic Separation and Positive Selection of Murine Neuronal Cultures

Magnetic Separation and Positive Selection of Murine Neuronal Cultures (Video) | JoVE 

小鼠神经元培养物的磁分离和正向选择