Differentiation of Human Mesenchymal Stem Cells from Wharton’s Jelly Towards Neural Stem Cells Using a Feasible and Repeatable Protocol

Abstract

The transplantation of neural stem cells (NSCs) capable of regenerating to the cells of the central nervous system (CNS) is a promising strategy in the treatment of CNS diseases and injury. As previous studies have highlighted mesenchymal stem cells (MSCs) as a source of NSCs, this study aimed to develop a feasible, efficient, and reproducible method for the neural induction of MSCs isolated from Wharton's jelly (hWJ-MSCs). We induced neural differentiation in a monolayer culture using epidermal growth factor, basic fibroblast growth factor, N2, and B27 supplements. This resulted in a homogenous population of proliferating cells that expressed certain neural markers at both the protein and mRNA levels. Flow cytometry and immunocytochemistry confirmed the expression of neural markers: nestin, sex-determining region Y (SRY) box 1 and 2 (SOX1 and SOX2), microtubule-associated protein 2 (MAP2), and glial fibrillary acidic protein (GFAP). The qRT-PCR analysis revealed significantly enhanced expression of nestin and MAP2 in differentiated cells. This study confirms that it is possible to generate NSCs-like cells from hWJ-MSCs in a 2D culture using a practical method. However, the therapeutic effectiveness of such differentiated cells should be extended to confirm the terminal differentiation ability and electrophysiological properties of neurons derived from them.

Phenotypical Characterization and Neurogenic Differentiation of Rabbit Adipose Tissue-Derived Mesenchymal Stem Cells

“…There is a wide range of neuronal markers dependent on the neuronal lineage being investigated, including glial fibrillary acidic protein (GFAP), microtubule-associated protein 2 (MAP2), nestin, neuron-specific enolase (ENO2) or β-III-tubulin [ 26 , 27 ]. The expression of specific neuronal markers (ENO2, MAP2) was confirmed in the present work in all types of rabbit MSCs (AT-MSCs, BM-MSCs and AF-MSCs) as well as in hAT-MSCs, similarly as in other studies on human MSCs from different sources [ 70 , 71 ]. Based on our results, which correspond to the previously reported findings [ 72 , 73 , 74 ], we can state that stem cells isolated from the adipose tissue show several advantages compared to the bone marrow.…”

Evaluation of Culture Morphology of Neuronally Transdifferentiated Wharton’s Jelly Derived Mesenchymal Stem Cells

“…This characteristic morphology was Evaluation of Culture Morphology of Neuronally Transdifferentiated Wharton's Jelly Derived Mesenchymal Stem Cells in accordance with Lija et al (2019), who succeeded in isolating and culturing neural stem cells from ovine fetal cerebral cortex. Similar results with regard to culture morphology were observed in neuronally transdifferentiated human WJ-MSCs, adipocyte derived MSCs and bone marrow derived MSCs (Kruminis et al, 2020;Zheng et al, 2017).…”

Significant transcriptomic changes are associated with differentiation of bone marrow-derived mesenchymal stem cells into neural progenitor-like cells in the presence of bFGF and EGF

“…Therapeutically, it may be more effective, however, to transplant endogenous neural stem cells or primary NPCs in neurological disorders but obtaining these endogenous stem or progenitor cells are extremely difficult in addition to ethical restrictions in their extraction. Hence, it is practical to use the MSCs-derived NPCs in neurodegenerative disorders [ 17 , 18 ]. Therefore, it is will be therapeutically more effective to enhance the differentiation of MSCs into NPCs.…”