脑皮质细胞分离培养 Culture of Dissociated Cortical Neurons
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脑皮质细胞分离培养 Culture of Dissociated Cortical Neurons
	1. Use 7 to 8 day chicken embryos
	
	2. Dissect forebrain in Ham's F12 media, peel off all meninges and then separate cortex from basal ganglia
	
	3. Dissociate neurons by incubating with 0.5% trypsin for 20 minutes at 37°C
	
	4. Wash tissue with fetal calf serum for 5 minutes to end trypsinization
	
	5. Dissociate tissue by gentle trituration with a flamed pipet
	
	6. Wash 2x in F12 medium, transfer to Neurobasal Medium (NBM, Gibco)
	
	7. Plate onto pretreated (poly-dl -ornithine or poly-l -lysine) culture dishes containing NBM or F12+ medium.
	
	8. Culture for 2 to 3 days
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