Alaylıoğlu Merve, Keskin Ebru, Yediel Büşra Şengül, Dursun Erdinç, Ak Duygu Gezen
Brain and Neurodegenerative Disorders Research Laboratories, Department of Neuroscience, Institute of Neurological Sciences, Istanbul University-Cerrahpasa, Istanbul, Turkey.
Department of Medical Biology, Cerrahpasa Faculty of Medicine, Istanbul University-Cerrahpasa, Istanbul, Turkey.
Noro Psikiyatr Ars. 2024 Aug 19;67(3):208-212. doi: 10.29399/npa.28510. eCollection 2024.
Human neuroblastoma cell line SH-SY5Y is a frequently used experimental cellular model in a variety of neuropsychiatric and neurodegenerative disorders. It is crucial to use a culture protocol that supports the fully differentiation of SH-SY5Y into neuron-like phenotype for the consistency of the results with neurons in vivo. However, a standardized neuronal differentiation protocol for SH-SY5Y cells still does not exist. Numerous differentiation methods have been proposed in the literature, yet SH-SY5Y cells with stronger neuronal characteristics and a more favorable environment for these differentiated cells are required in order to best representation of neurons. Therefore, in the study, we aimed to establish a more successful differentiation protocol for SH-SY5Y cells based on the primary neuron culture technique, which neuronal maturation is very well defined.
In the study, we rearranged previous SH-SY5Y differentiation protocols, combined them with our primary neuron culture protocol and created a robust and reproducible protocol for differentiation of SH-SY5Y.
Our proposed "retinoic acid+brain-derived neurotrophic factor (RA+BDNF)-induced 7 days differentiation (conalbumin- on day 4) protocol provided well developed neurites, adequate expression and localization of neuronal and synaptic markers resembling mature neurons.
The differentiation protocol we present can enable researchers to obtain satisfactory and properly differentiated SH-SY5Y cells in each independent experiment, achieving the closest possible in vivo results.
人神经母细胞瘤细胞系SH-SY5Y是多种神经精神疾病和神经退行性疾病中常用的实验细胞模型。为了使结果与体内神经元保持一致，使用一种支持SH-SY5Y细胞完全分化为神经元样表型的培养方案至关重要。然而，目前仍不存在针对SH-SY5Y细胞的标准化神经元分化方案。文献中已经提出了许多分化方法，但为了更好地模拟神经元，仍需要具有更强神经元特征且对这些分化细胞有更有利环境的SH-SY5Y细胞。因此，在本研究中，我们旨在基于原代神经元培养技术建立一种更成功的SH-SY5Y细胞分化方案，该技术中神经元的成熟过程已被明确界定。
在本研究中，我们重新整理了之前的SH-SY5Y分化方案，将它们与我们的原代神经元培养方案相结合，创建了一种稳健且可重复的SH-SY5Y分化方案。
我们提出的“视黄酸+脑源性神经营养因子（RA+BDNF）诱导7天分化（第4天添加伴清蛋白）方案”产生了发育良好的神经突，神经元和突触标记物的表达和定位充足，类似于成熟神经元。
我们提出的分化方案能够使研究人员在每个独立实验中获得令人满意且分化良好的SH-SY5Y细胞，取得尽可能接近体内实验的结果。
