In-vitro culture method for rat olfactory bulb neurons

By rolling peeling method and papaya protease mixed digestive technology with DNASE I enzymes, combined with the Neurobasal basic medium and B27 additives, the problem of incomplete digestion and divestitudes of the olfactory neuron in vitro of the olfactory ball has been solved. And good growth environment.

CN119351332AActive Publication Date: 2025-01-24KUNMING MEDICAL UNIVERSITY
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Patent Information

Application Number
CN202411932182.6
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2024-12-26
Publication Date
2025-01-24
Estimated Expiration
2044-12-26

AI Technical Summary

Technical Problem

In the prior art, incomplete digestion or excessive digestion of olfactory bulb neurons leads to cell destruction, affecting neuronal growth and observation, and severe loss during meninges peeling, which increases culture cost and errors.

Method used

The olfactory bulb meninges were peeled off using the rolling dissection method, and mixed digestion with papain and DNase I enzymes was used to gently digest and prevent cell adhesion. Neurobasal basal medium and B27 additives were used to improve neuron survival activity, and the medium change time was extended to promote neuron adhesion. wall growth.

Benefits of technology

The efficient and stable in vitro cultivation of the olfactory ball neurons is achieved, the cell purity and activity is improved, the cell block is reduced, the survival time of the neuron is prolonged, and the experimental cost and time are reduced.

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Abstract

The invention discloses an in-vitro culture method for rat olfactory bulb neurons, and belongs to the field of cell culture. The method comprises the following steps: stripping meningeal membranes of rats by using a rolling stripping method to obtain olfactory bulbs, washing, cutting into pieces, digesting by using DNaseI enzyme and papain, centrifuging, resuspending, filtering, and performing adherent culture to obtain the rat olfactory bulb neurons. The method is simple to operate and good in repeatability, time and experiment cost are saved, and the cultured olfactory bulb neurons are uniform in distribution, good in stability, high in activity and long in in-vitro survival time.
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Citation Information

Patent Citations

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  • Method of making primary culture of olfactory neurons

    US5308763A