Immunological isolation and characterization of neuronal progenitors from human dental pulp: A laboratory-based investigation.

Autor: McMillan HP; School of Medicine, Dentistry and Biomedical Sciences, Queens University Belfast, Northern Ireland, UK., Lundy FT; School of Medicine, Dentistry and Biomedical Sciences, Queens University Belfast, Northern Ireland, UK., Dunne OM; School of Medicine, Dentistry and Biomedical Sciences, Queens University Belfast, Northern Ireland, UK., McLoughlin KJ; School of Medicine, Dentistry and Biomedical Sciences, Queens University Belfast, Northern Ireland, UK., About I; Aix Marseille University, CNRS, Institute of Movement Sciences, Marseille, France., Curtis TM; School of Medicine, Dentistry and Biomedical Sciences, Queens University Belfast, Northern Ireland, UK., El Karim I; School of Medicine, Dentistry and Biomedical Sciences, Queens University Belfast, Northern Ireland, UK.
Jazyk: angličtina
Zdroj: International endodontic journal [Int Endod J] 2024 Aug; Vol. 57 (8), pp. 1136-1146. Date of Electronic Publication: 2024 May 07.
DOI: 10.1111/iej.14077
Abstrakt: Aims: Dental pulp stem cells (DPSCs) contain a population of stem cells with a broad range of differentiation potentials, as well as more lineage-committed progenitors. Such heterogeneity is a significant obstacle to experimental and clinical applications. The aim of this study is to isolate and characterize a homogenous neuronal progenitor cell population from human DPSCs.
Methodology: Polysialylated-neural cell adhesion molecule (PSA-NCAM+) neural progenitors were isolated from the dental pulp of three independent donors using magnetic-activated cell sorting (MACS) technology. Immunofluorescent staining with a panel of neural and non-neural markers was used to characterize the magnetically isolated PSA-NCAM+ fraction. PSA-NCAM+ cells were then cultured in Neurobasal A supplemented with neurotrophic factors: dibutyryl cyclic-AMP, neurotrophin-3, B27 and N2 supplements to induce neuronal differentiation. Both PSA-NCAM+ and differentiated PSA-NCAM+ cells were used in Ca 2+ imaging studies to assess the functionality of P2X3 receptors as well as membrane depolarization.
Results: PSA-NCAM+ neural progenitors were isolated from a heterogeneous population of hDPSCs using magnetic-activated cell sorting and anti-PSA-NCAM MicroBeads. Flow cytometry analysis demonstrated that immunomagnetic sorting significantly increased the purity of PSA-NCAM+ cells. Immunofluorescent staining revealed expression of pan-neuronal and mature neuronal markers, PGP9.5 and MAP2, respectively, as well as weak expression of the mature sensory markers, peripherin and islet1. ATP-induced response was mediated predominately by P2X3 receptors in both undifferentiated and differentiated cells, with a greater magnitude observed in the latter. In addition, membrane depolarizations were also detected in cells before and after differentiation when loaded with fast-voltage-responding fluorescent molecule, FluoVolt™ in response to potassium chloride. Interestingly, only differentiated PSA-NCAM+ cells were capable of spontaneous membrane oscillations.
Conclusions: In summary, DPSCs contain a population of neuronal progenitors with enhanced neural differentiation and functional neural-like properties that can be effectively isolated with magnetic-activated cell sorting (MACS).
(© 2024 The Authors. International Endodontic Journal published by John Wiley & Sons Ltd on behalf of British Endodontic Society.)
Databáze: MEDLINE
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