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Role of cerebrospinal fluid in differentiation of human dental pulp stem cells into neuron-like cells

Anatomy & Cell Biology 2020년 53권 3호 p.292 ~ 300
Goudarzi Ghazaleh, Hamidabadi Hatef Ghasemi, Bojnordi Maryam Nazm, Hedayatpour Azim, Niapour Ali, Zahiri Maria, Absalan Forouzan, Darabi Shahram,
소속 상세정보
 ( Goudarzi Ghazaleh ) - Mazandaran University of Medical Sciences Faculty of Medicine Department of Anatomy and Cell Biology
 ( Hamidabadi Hatef Ghasemi ) - Mazandaran University of Medical Sciences Faculty of Medicine Department of Anatomy and Cell Biology
 ( Bojnordi Maryam Nazm ) - Mazandaran University of Medical Sciences Faculty of Medicine Department of Anatomy and Cell Biology
 ( Hedayatpour Azim ) - Tehran University of Medical Sciences School of Medicine Department of Anatomy
 ( Niapour Ali ) - Ardabil University of Medical Sciences School of Medicine Department of Anatomical Sciences
 ( Zahiri Maria ) - Bushehr University of Medical Sciences School of Medical Sciences Department of Anatomical Sciences
 ( Absalan Forouzan ) - Abadan School of Medical Sciences
 ( Darabi Shahram ) - Qazvin University of Medical Sciences Cellular and Molecular Research Center

Abstract


Human dental pulp stem cells (hDPSCs) could be differentiated into neuron like-cells under particular microenvironments. It has been reported that a wide range of factors, presented in cerebrospinal fluid (CSF), playing part in neuronal differentiation during embryonic stages, we herein introduce a novel culture media complex to differentiate hDPSCs into neuron-like cells. The hDPSCs were initially isolated and characterized. The CSF was prepared from the Cisterna magna of 19-day-old Wistar rat embryos, embryonic cerebrospinal fluid (E-CSF). The hDPSCs were treated by 5% E-CSF for 2 days, then neurospheres were cultured in DMEM/F12 supplemented with 10-6 μm retinoic acid (RA), glial-derived neurotrophic factor and brain-derived neurotrophic factor for 6 days. The cells which were cultured in basic culture medium were considered as control group. Morphology of differentiated cells as well as process elongation were examined by an inverted microscope. In addition, the neural differentiation markers (Nestin and MAP2) were studied employing immunocytochemistry. Neuronal-like processes appeared 8 days after treatment. Neural progenitor marker (Nestin) and a mature neural marker (MAP2) were expressed in treated group. Moreover Nissl bodies were found in the cytoplasm of treated group. Taking these together, we have designed a simple protocol for generating neuron-like cells using CSF from the hDPSCs, applicable for cell therapy in several neurodegenerative disorders including Alzheimer’s disease.

키워드

Human dental pulp stem cells; Cerebrospinal fluid; Cell transdifferentiation; Alzheimer

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