SOCS7-Derived BC-Box Motif Peptide Mediated Cholinergic Differentiation of Human Adipose-Derived Mesenchymal Stem Cells
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Published:2023-02-01
Issue:3
Volume:24
Page:2786
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ISSN:1422-0067
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Container-title:International Journal of Molecular Sciences
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language:en
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Short-container-title:IJMS
Author:
Kanno Hiroshi123ORCID, Matsumoto Shutaro12, Yoshizumi Tetsuya134, Nakahara Kimihiro3, Shinonaga Masamichi3, Kubo Atsuhiko5, Fujii Satoshi2, Ishizuka Yasuyuki6ORCID, Tanaka Masaki7, Ichihashi Masamitsu7ORCID, Murata Hidetoshi4
Affiliation:
1. Department of Neurosurgery, Yokohama City University, Yokohama 236-0004, Japan 2. Department of Neurosurgery, Asahi Hospital, Tokyo 121-0078, Japan 3. Department of Neurosurgery, International University of Health and Welfare Atami Hospital, Atami 413-0012, Japan 4. Department of Neurosurgery, St. Marianna Medical University of Medicine, Kawasaki 216-8511, Japan 5. Nerve Care Clinic, Yokosuka 238-0012, Japan 6. BioFuture Technologies Ltd., Tokyo 105-0014, Japan 7. BTR Arts Ginza Clinic, Tokyo 105-0004, Japan
Abstract
Adipose-derived mesenchymal stem cells (ADMSCs) are a type of pluripotent somatic stem cells that differentiate into various cell types such as osteoblast, chondrocyte, and neuronal cells. ADMSCs as donor cells are used to produce regenerative medicines at hospitals and clinics. However, it has not been reported that ADMSCs were differentiated to a specific type of neuron with a peptide. Here, we report that ADMSCs differentiate to the cholinergic phenotype of neurons by the SOCS7-derived BC-box motif peptide. At operations for patients with neurological disorders, a small amount of subcutaneous fat was obtained. Two weeks later, adipose-derived mesenchymal stem cells (ADMSCs) were isolated and cultured for a further 1 to 2 weeks. Flow cytometry analysis for characterization of ADMSCs was performed with CD73, CD90, and CD105 as positive markers, and CD14, CD31, and CD56 as negative markers. The results showed that cultured cells were compatible with ADMSCs. Immunocytochemical studies showed naïve ADMSCs immunopositive for p75NTR, RET, nestin, keratin, neurofilament-M, and smooth muscle actin. ADMSCs were suggested to be pluripotent stem cells. A peptide corresponding to the amino-acid sequence of BC-box motif derived from SOCS7 protein was added to the medium at a concentration of 2 μM. Three days later, immunocytochemistry analysis, Western blot analysis, ubiquitination assay, and electrophysiological analysis with patch cramp were performed. Immunostaining revealed the expression of neurofilament H (NFH), choline acetyltransferase (ChAT), and tyrosine hydroxylase (TH). In addition, Western blot analysis showed an increase in the expression of NFH, ChAT, and TH, and the expression of ChAT was more distinct than TH. Immunoprecipitation with JAK2 showed an increase in the expression of ubiquitin. Electrophysiological analysis showed a large holding potential at the recorded cells through path electrodes. The BC-box motif peptide derived from SOCS7 promoted the cholinergic differentiation of ADMSCs. This novel method will contribute to research as well as regenerative medicine for cholinergic neuron diseases.
Funder
Ministry of Education, Culture, Sports, Science, and Technology of Japan
Subject
Inorganic Chemistry,Organic Chemistry,Physical and Theoretical Chemistry,Computer Science Applications,Spectroscopy,Molecular Biology,General Medicine,Catalysis
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