Calcium: A novel and efficient inducer of differentiation of adipose‐derived stem cells into neuron‐like cells

• Published in: Journal of cellular physiology 2018-11, Vol.233 (11), p.8940-8951
• Main Authors: Goudarzi, Farjam, Tayebinia, Heidar, Karimi, Jamshid, Habibitabar, Elahe, Khodadadi, Iraj
• Format: Article
• Language: English
• Subjects: adipose tissue; Biomarkers; Calcium; Calcium mobilization; Cell cycle; cell differentiation; Cell proliferation; Differentiation; Doublecortin protein; Gene expression; Glial fibrillary acidic protein; Immunofluorescence; Insulin; Mesenchymal stem cells; mesenchymal stromal cells; Myelin; Myelin basic protein; Neuronal-glial interactions; neurons; Phosphopyruvate hydratase; Polymerase chain reaction; Proteins; Stem cells; Synapsin I
• ISSN: 0021-9541; 1097-4652
• DOI: 10.1002/jcp.26826

This study comparatively investigated the effectiveness of calcium and other well‐known inducers such as isobutylmethylxanthine (IBMX) and insulin in differentiating human adipose‐derived stem cells (ADSCs) into neuronal‐like cells. ADSCs were immunophenotyped and differentiated into neuron‐like cells with different combinations of calcium, IBMX, and insulin. Calcium mobilization across the membrane was determined. Differentiated cells were characterized by cell cycle profiling, staining of Nissl bodies, detecting the gene expression level of markers such as neuronal nuclear antigen (NeuN), microtubule associated protein 2 (MAP2), neuron‐specific enolase (NSE), doublecortin, synapsin I, glial fibrillary acidic protein (GFAP), and myelin basic protein (MBP) by quantitative real‐time polymerase chain reaction (quantitative real‐time polymerase chain reaction (qRT‐PCR) and protein level by the immunofluorescence technique. Treatment with Ca + IBMX + Ins induced neuronal appearance and projection of neurite‐like processes in the cells, accompanied with inhibition of proliferation and halt in the cell cycle. A significantly higher expression of MBP, GFAP, NeuN, NSE, synapsin 1, doublecortin, and MAP2 was detected in differentiated cells, confirming the advantages of Ca + IBMX + Ins to the other combinations of inducers. Here, we showed an efficient protocol for neuronal differentiation of ADSCs, and calcium fostered differentiation by augmenting the number of neuron‐like cells and instantaneous increase in the expression of neuronal markers. The insufficiency of current protocols in differentiating mesenchymal stem cells into a pure neuronal phenotype has limited the applicability of these cells in therapeutic approaches. The current study showed that calcium fostered differentiation by augmenting the number of neuron‐like cells and instantaneous increase in the expression of neuronal markers, and therefore the mixture of Ca + IBMX + Ins can be introduced as a novel and potent differentiation reagent.