|Jezik rada|| engleski |
|Naslov (engleski)||An improved method for growing neurons: Comparison with standard protocols |
|Autor||Pozzi, Diletta |
|Sažetak rada (engleski)|| |
Background: Since different culturing parameters – such as media composition or cell density – lead to different experimental results, it is important to define the protocol used for neuronal cultures. The vital role of astrocytes in maintaining homeostasis of neurons – both in vivo and in vitro – is well established: the majority of improved culturing conditions for primary dissociated neuronal cultures rely on astrocytes.
New method: Our culturing protocol is based on a novel serum-free preparation of astrocyte – conditioned medium (ACM). We compared the proposed ACM culturing method with other two commonly used methods Neurobasal/B27- and FBS- based media. We performed morphometric characterization by immunocytochemistry and functional analysis by calcium imaging for all three culture methods at 1, 7,14 and 60 days in vitro (DIV).
Results: ACM-based cultures gave the best results for all tested criteria, i.e. growth cone’s size and shape, neuronal outgrowth and branching, network activity and synchronization, maturation and long-term survival. The differences were more pronounced when compared with FBS-based medium. Neurobasal/B27cultures were comparable to ACM for young cultures (DIV1), but not for culturing times longer than DIV7.
Comparison with existing method(s): ACM-based cultures showed more robust neuronal outgrowth at DIV1. At DIV7 and 60, the activity of neuronal network grown in ACM had a more vigorous spontaneous electrical activity and a higher degree of synchronization.
Conclusions: We propose our ACM-based culture protocol as an improved and more suitable method for both short- and long-term neuronal cultures.
|Ključne riječi (engleski)||Neuronal culture, Astrocyte, B27 Supplement, Fetal bovine serum, Neuronal branching, Calcium signaling |
|Vrsta rada||znanstveni rad - izvorni znanstveni rad |
|Status objave rada||objavljen |
|Vrsta recenzije||recenziran - međunarodna recenzija |
|Naslov časopisa||Journal of Neuroscience Methods |
|Brojčani podaci||2017, Vol. 120, str. 1-10 |
|Datum||datum objave publikacije: 15.03.2017. |
|Oznaka DOI||https://doi.org/10.1016/j.jneumeth.2017.01.013 |
|URL rada||https://www.sciencedirect.com/science/article/pii/S0165027017300195?via%3Dihub |
|Znanstveno područje||INTERDISCIPLINARNA PODRUČJA ZNANOSTI |
Biotehnologija u biomedicini (prirodno područje, biomedicina i zdravstvo, biotehničko područje)
|Ustanova||Sveučilište u Rijeci |
(Odjel za biotehnologiju)