Transformation of SH-SY5Y cell line into neuron-like cells: Investigation of electrophysiological and biomechanical changes

• Published in: Neuroscience letters 2021-02, Vol.745, p.135628, Article 135628
• Main Authors: Şahin, Meryem, Öncü, Gül, Yılmaz, Mustafa Alper, Özkan, Doğuş, Saybaşılı, Hale
• Format: Article
• Language: English
• Subjects: Atomic force microscopy; Biomechanical Phenomena - drug effects; Biomechanical Phenomena - physiology; Brain-Derived Neurotrophic Factor - pharmacology; Cell Differentiation - drug effects; Cell Differentiation - physiology; Cell Line, Tumor; Cell Proliferation - drug effects; Cell Proliferation - physiology; Cell Transformation, Neoplastic - drug effects; Cell Transformation, Neoplastic - metabolism; Cell Transformation, Neoplastic - pathology; Electrophysiological Phenomena - drug effects; Electrophysiological Phenomena - physiology; Electrophysiology; Humans; Microscopy, Atomic Force - methods; Neuroblastoma; Neurons - drug effects; Neurons - metabolism; Neurons - pathology; SH-SY5Y; Tretinoin - pharmacology
• ISSN: 0304-3940; 1872-7972
• DOI: 10.1016/j.neulet.2021.135628

•Electrophysiology of undifferentiated and differentiated SH-SY5Y cells was studied.•Elasticity of SH-SY5Y cells was examined by atomic force microscopy.•Undifferentiated SH-SY5Y cells had large outward K+ currents and low elasticity.•Na+ currents were observed in cells differentiated with retinoic acid and BDNF. SH-SY5Y human neuroblastoma cells are commonly used as neuronal models. Here, we examined different aspects of SH-SY5Y cell differentiation. Various differentiation protocols have been proposed previously, including treatments with retinoic acid, brain-derived neurotrophic factor (BDNF), cholesterol and oestradiol. We examined undifferentiated SH-SY5Y cells (UNDIFF); cells differentiated by the treatment with retinoic acid (RA); retinoic acid + BDNF (RB); and retinoic acid + BDNF + cholesterol + oestradiol (RBCE). We performed whole-cell patch-clamp recordings from these cells and nanomechanically characterised them by using atomic force microscopy (AFM). Our results indicated that Na+ currents become most pronounced in the differentiated RB cells, whereas UNDIFF SH-SY5Y cells had significantly larger K+ currents, which is a characteristic feature of cancer cells. AFM observations of these two groups showed that Young’s moduli of SH-SY5Y cells increased threefold with differentiation. Furthermore, we showed a direct relationship between Na+ channel activity and elasticity in these cells. We conclude that SH-SY5Y human neuroblastoma cells should be used as a neuronal model only when they are differentiated by the treatment with retinoic acid and BDNF.