Clonal Heterogeneity in the Neuronal and Glial Differentiation of Dental Pulp Stem/Progenitor Cells

Abstract

Cellular heterogeneity presents an important challenge to the development of cell-based therapies where there is a fundamental requirement for predictable and reproducible outcomes. Transplanted Dental Pulp Stem/Progenitor Cells (DPSCs) have demonstrated early promise in experimental models of spinal cord injury and stroke, despite limited evidence of neuronal and glial-like differentiation after transplantation. Here, we report, for the first time, on the ability of single cell-derived clonal cultures of murine DPSCs to differentiate in vitro into immature neuronal-like and oligodendrocyte-like cells. Importantly, only DPSC clones with high nestin mRNA expression levels were found to successfully differentiate into Map2 and NF-positive neuronal-like cells. Neuronally differentiated DPSCs possessed a membrane capacitance comparable with primary cultured striatal neurons and small inward voltage-activated K+ but not outward Na+ currents were recorded suggesting a functionally immature phenotype. Similarly, only high nestin-expressing clones demonstrated the ability to adopt Olig1, Olig2, and MBP-positive immature oligodendrocyte-like phenotype. Together, these results demonstrate that appropriate markers may be used to provide an early indication of the suitability of a cell population for purposes where differentiation into a specific lineage may be beneficial and highlight that further understanding of heterogeneity within mixed cellular populations is required.

Andrographolide Promotes Neural Differentiation of Rat Adipose Tissue-Derived Stromal Cells through Wnt/β-Catenin Signaling Pathway

“…Compared to control cultures maintained in the neural differentiation medium alone, ANDRO-treated ADSCs exhibited enhanced expression of the mature neuronal marker MAP2 [ 33 ] and β -tubulin III, which is an important structural protein for early neurons. While both ANDRO-treated and control cultures appeared to exhibit similar marker expression levels as assessed by immunofluorescence analysis, protein expression as estimated by Western blotting was significantly higher in cultures treated with 10 μ M ANDRO.…”

Cellular and Molecular Mechanisms Mediated by recPrPC Involved in the Neuronal Differentiation Process of Mesenchymal Stem Cells

“…In a previous work, we used human dental pulp stem cell (hDPSCs) as the experimental model to develop an in vitro system used to study proliferation and differentiation process [22]. hDPSCs represent a kind of adult mesenchymal stem cells that show the ability to differentiate in vitro in several multilineages such as odontoblasts, osteoblasts, chondrocytes, adipocytes and neurons [23,24,25,26,27,28,29,30]. The fact that hDPSCs can express basally PrP C [21] and show the ability to differentiate into neuronal-like cells [31] or dopaminergic neuron-like cells [32], prompt it to be considered as a cellular model for the study of neurodegenerative diseases, such as Alzheimer, Parkinson and Huntington diseases [22,33,34].…”

Role of Prion protein-EGFR multimolecular complex during neuronal differentiation of human dental pulp-derived stem cells

“…CD44, CD90, CD105, STRO-1) and are negative for hematopoietic markers (CD14, CD19), but capable of in vitro differentiation into odontoblasts, osteoblasts, chondrocytes, adipocytes and neurons [8][9][10]. Several works have shown that hDPSCs represent a highly heterogeneous population with distinct clones and differences in proliferative and differentiating capacity [11,12]. In particular, hDPSCs show the ability to differentiate into neuronallike cells [13] or dopaminergic neuron-like cells [14].…”