Cargando…

Comparative analysis of three different protocols for cholinergic neuron differentiation in vitro using mesenchymal stem cells from human dental pulp

A decrease in the activity of choline acetyltransferase, the enzyme responsible for acetylcholine synthesis in the cholinergic neurons cause neurological disorders involving a decline in cognitive abilities, such as Alzheimer’s disease. Mesenchymal stem cells (MSCs) can be used as an efficient thera...

Descripción completa

Detalles Bibliográficos
Autores principales: Kang, Young-Hoon, Shivakumar, Sharath Belame, Son, Young-Bum, Bharti, Dinesh, Jang, Si-Jung, Heo, Kang-Sun, Park, Won-Uk, Byun, June-Ho, Park, Bong-Wook, Rho, Gyu-Jin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6711138/
https://www.ncbi.nlm.nih.gov/pubmed/31489249
http://dx.doi.org/10.1080/19768354.2019.1626280
_version_ 1783446466898952192
author Kang, Young-Hoon
Shivakumar, Sharath Belame
Son, Young-Bum
Bharti, Dinesh
Jang, Si-Jung
Heo, Kang-Sun
Park, Won-Uk
Byun, June-Ho
Park, Bong-Wook
Rho, Gyu-Jin
author_facet Kang, Young-Hoon
Shivakumar, Sharath Belame
Son, Young-Bum
Bharti, Dinesh
Jang, Si-Jung
Heo, Kang-Sun
Park, Won-Uk
Byun, June-Ho
Park, Bong-Wook
Rho, Gyu-Jin
author_sort Kang, Young-Hoon
collection PubMed
description A decrease in the activity of choline acetyltransferase, the enzyme responsible for acetylcholine synthesis in the cholinergic neurons cause neurological disorders involving a decline in cognitive abilities, such as Alzheimer’s disease. Mesenchymal stem cells (MSCs) can be used as an efficient therapeutic agents due to their neuronal differentiation potential. Different source derived MSCs may have different differentiation potential under different inductions. Various in vitro protocols have been developed to differentiate MSCs into specific neurons but the comparative effect of different protocols utilizing same source derived MSCs, is not known. To address this issue, dental pulp derived MSCs (DPSCs) were differentiated into cholinergic neurons using three different protocols. In protocol I, DPSCs were pre-induced with serum-free ADMEM containing 1 mM of β-mercaptoethanol for 24 h and then incubated with 100 ng/ml nerve growth factor (NGF) for 6 days. Under protocol II, DPSCs were cultured in serum-free ADMEM containing 15 µg/ml of D609 (tricyclodecan-9-yl-xanthogenate) for 4 days. Under protocol III, the DPSCs were cultured in serum-free ADMEM containing 10 ng/ml of basic fibroblast growth factor (bFGF), 50 µM of forskolin, 250 ng/ml of sonic hedgehog (SHH), and 0.5 µM of retinoic acid (RA) for 7 days. The DPSCs were successfully trans-differentiated under all the protocols, exhibited neuron-like morphologies with upregulated cholinergic neuron-specific markers such as ChAT, HB9, ISL1, BETA-3, and MAP2 both at mRNA and protein levels in comparison to untreated cells. However, protocol III-induced cells showed the highest expression of the cholinergic markers and secreted the highest level of acetylcholine.
format Online
Article
Text
id pubmed-6711138
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher Taylor & Francis
record_format MEDLINE/PubMed
spelling pubmed-67111382019-09-05 Comparative analysis of three different protocols for cholinergic neuron differentiation in vitro using mesenchymal stem cells from human dental pulp Kang, Young-Hoon Shivakumar, Sharath Belame Son, Young-Bum Bharti, Dinesh Jang, Si-Jung Heo, Kang-Sun Park, Won-Uk Byun, June-Ho Park, Bong-Wook Rho, Gyu-Jin Anim Cells Syst (Seoul) Neurobiology & Physiology A decrease in the activity of choline acetyltransferase, the enzyme responsible for acetylcholine synthesis in the cholinergic neurons cause neurological disorders involving a decline in cognitive abilities, such as Alzheimer’s disease. Mesenchymal stem cells (MSCs) can be used as an efficient therapeutic agents due to their neuronal differentiation potential. Different source derived MSCs may have different differentiation potential under different inductions. Various in vitro protocols have been developed to differentiate MSCs into specific neurons but the comparative effect of different protocols utilizing same source derived MSCs, is not known. To address this issue, dental pulp derived MSCs (DPSCs) were differentiated into cholinergic neurons using three different protocols. In protocol I, DPSCs were pre-induced with serum-free ADMEM containing 1 mM of β-mercaptoethanol for 24 h and then incubated with 100 ng/ml nerve growth factor (NGF) for 6 days. Under protocol II, DPSCs were cultured in serum-free ADMEM containing 15 µg/ml of D609 (tricyclodecan-9-yl-xanthogenate) for 4 days. Under protocol III, the DPSCs were cultured in serum-free ADMEM containing 10 ng/ml of basic fibroblast growth factor (bFGF), 50 µM of forskolin, 250 ng/ml of sonic hedgehog (SHH), and 0.5 µM of retinoic acid (RA) for 7 days. The DPSCs were successfully trans-differentiated under all the protocols, exhibited neuron-like morphologies with upregulated cholinergic neuron-specific markers such as ChAT, HB9, ISL1, BETA-3, and MAP2 both at mRNA and protein levels in comparison to untreated cells. However, protocol III-induced cells showed the highest expression of the cholinergic markers and secreted the highest level of acetylcholine. Taylor & Francis 2019-06-10 /pmc/articles/PMC6711138/ /pubmed/31489249 http://dx.doi.org/10.1080/19768354.2019.1626280 Text en © 2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Neurobiology & Physiology
Kang, Young-Hoon
Shivakumar, Sharath Belame
Son, Young-Bum
Bharti, Dinesh
Jang, Si-Jung
Heo, Kang-Sun
Park, Won-Uk
Byun, June-Ho
Park, Bong-Wook
Rho, Gyu-Jin
Comparative analysis of three different protocols for cholinergic neuron differentiation in vitro using mesenchymal stem cells from human dental pulp
title Comparative analysis of three different protocols for cholinergic neuron differentiation in vitro using mesenchymal stem cells from human dental pulp
title_full Comparative analysis of three different protocols for cholinergic neuron differentiation in vitro using mesenchymal stem cells from human dental pulp
title_fullStr Comparative analysis of three different protocols for cholinergic neuron differentiation in vitro using mesenchymal stem cells from human dental pulp
title_full_unstemmed Comparative analysis of three different protocols for cholinergic neuron differentiation in vitro using mesenchymal stem cells from human dental pulp
title_short Comparative analysis of three different protocols for cholinergic neuron differentiation in vitro using mesenchymal stem cells from human dental pulp
title_sort comparative analysis of three different protocols for cholinergic neuron differentiation in vitro using mesenchymal stem cells from human dental pulp
topic Neurobiology & Physiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6711138/
https://www.ncbi.nlm.nih.gov/pubmed/31489249
http://dx.doi.org/10.1080/19768354.2019.1626280
work_keys_str_mv AT kangyounghoon comparativeanalysisofthreedifferentprotocolsforcholinergicneurondifferentiationinvitrousingmesenchymalstemcellsfromhumandentalpulp
AT shivakumarsharathbelame comparativeanalysisofthreedifferentprotocolsforcholinergicneurondifferentiationinvitrousingmesenchymalstemcellsfromhumandentalpulp
AT sonyoungbum comparativeanalysisofthreedifferentprotocolsforcholinergicneurondifferentiationinvitrousingmesenchymalstemcellsfromhumandentalpulp
AT bhartidinesh comparativeanalysisofthreedifferentprotocolsforcholinergicneurondifferentiationinvitrousingmesenchymalstemcellsfromhumandentalpulp
AT jangsijung comparativeanalysisofthreedifferentprotocolsforcholinergicneurondifferentiationinvitrousingmesenchymalstemcellsfromhumandentalpulp
AT heokangsun comparativeanalysisofthreedifferentprotocolsforcholinergicneurondifferentiationinvitrousingmesenchymalstemcellsfromhumandentalpulp
AT parkwonuk comparativeanalysisofthreedifferentprotocolsforcholinergicneurondifferentiationinvitrousingmesenchymalstemcellsfromhumandentalpulp
AT byunjuneho comparativeanalysisofthreedifferentprotocolsforcholinergicneurondifferentiationinvitrousingmesenchymalstemcellsfromhumandentalpulp
AT parkbongwook comparativeanalysisofthreedifferentprotocolsforcholinergicneurondifferentiationinvitrousingmesenchymalstemcellsfromhumandentalpulp
AT rhogyujin comparativeanalysisofthreedifferentprotocolsforcholinergicneurondifferentiationinvitrousingmesenchymalstemcellsfromhumandentalpulp