Isolation and Culturing of Primary Neurons from Newborn Mouse Cortex Tissue

Year 2024, Volume: 13 Issue: 3, 98 - 104, 31.12.2024

Nese Ayşit

https://doi.org/10.47493/abantmedj.1479470

Abstract

Objective: This study aimed to establish a reliable protocol for obtaining healthy and long-lived cortical neurons from newborn mice, providing a valuable model for studying neuronal function.
Materials and Methods: Cortical regions of P0 mice were isolated and healthy neurons were obtained by enzymatic and mechanical dissociation. On the seventh day of incubation, the presence of neurons was detected by staining with neuron-specific antibodies. Transgenic mice expressing fluorescent proteins specific to neurons, glia and oligodendrocytes were also used for culture.
Results: Almost all the neurons had adhered to the petri dish bottom by the second hour of incubation. Most of the neurons were healthy and started to grow extension quickly.
Conclusion: Neuron cultures are an important tool in research and are invaluable for studying the behaviour of cells. Nanoparticles facilitate genetic manipulation of these cultures for various biotechnological applications. In particular, they have great potential in areas such as the delivery of genetic material into cells, drug delivery and targeted treatment methods. Such techniques have the potential to open up new avenues for the study and treatment of neurological diseases.

Keywords

Cortex, Newborn Mice, Cell Culture, Neuron Culture, Central Nervous System.

Project Number

The project did not provide any support for our work.

Yeni Doğan Farelerden İzole Edilen Korteks Dokusundan Primer Nöron Elde Edilmesi

Abstract

Amaç: Bu çalışma, yeni doğmuş farelerden sağlıklı ve uzun ömürlü kortikal nöronlar elde etmek için güvenilir bir protokol geliştirmeyi ve elde edilen nöronları kullanarak nöron fonksiyonlarını incelemeyi hedeflemiştir.
Gereç ve Yöntemler: P0 farelerin korteks bölgeleri izole edilerek enzimatik ve mekanik ayrıştırma uygulanarak sağlıklı nöronlar elde edildi. İnkübasyonun yedinci gününde nörona özgü antikorlar ile boyama yapılarak nöronların varlığı gösterildi. Ayrıca nöron, glia ve oligodendrositlere özgü floresan proteinleriifade eden transgenik farelerden de kültür yapıldı.
Bulgular: İnkübasyonun ikinci saatinde nöronların neredeyse tamamının kültür kabına yapıştığı gözlemlendi. Elde edilen nöronların büyük kısmının sağlıklı olduğu ve uzantılarının hızlıca büyümeye başladığı gözlemlendi.
Sonuç: Bu protokolü diğer protokollerden ayıran temel özellik, geliştirme sürecini destekleyecek hiçbir faktör veya serum kullanılmamasıdır. Nöron kültürleri, araştırmalarda önemli bir araç olarak kullanılır ve hücrelerin davranışlarını incelemek için oldukça muazzam olanak sağlar. Nanoparçacıklar, bu kültürler üzerinde çeşitli biyoteknolojik uygulamalar için genetik manipülasyonları kolaylaştırır. Özellikle, hücrelere genetik materyal taşıma, ilaç salınımı ve hedeflenmiş tedavi yöntemleri gibi alanlarda büyük potansiyele sahipler. Bu tür teknikler, nörolojik hastalıkların araştırılması ve tedavisi için yeni yollar açabilir. Ayrıca kısa sürede farklı amaçlar için tasarlanan nanoparçacık, biyoteknolojik araçlarla genetik manipülasyonlara olanak sağlar Geliştirilen protokol, bir nöronal gelişim, fizyolojik bir nöronal aktivite sergileyen tekrarlanabilir kültürler sağlar.

Keywords

Korteks, Yenidoğan Fare, Hücre Kültürü, Nöron Kültürü, Merkezi Sinir Sistemi.

Ethical Statement

This is an original study; the study design, data collection and analysis were carried out in accordance with the principles and rules of scientific ethics at all stages, including the presentation of information and the way in which I have handled all data and information not obtained within the framework of this study, that I have cited these sources and included them in the bibliography; all data used, that I have not altered them, and that the study complies with all the terms and conditions of the Abant Medical Journal. I declare that by accepting the terms and conditions I am fulfilling my ethical duties and responsibilities.

Supporting Institution

The project had no funding from any institution.

Project Number

The project did not provide any support for our work.

Thanks

Thank you to Istanbul Medipol University Experimental Animals Unit (MEDİTAM) for providing experimental animals.

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