Production of Neural Progenitors from Bone Marrow Mesenchymal Stem Cells

Maeda Mohammad,Ahmad Al-Joubory,Nahi Yaseen,Aesar A Ahmed,Rafal Abdullah,Ahmed Al-Shammari,Noah Mahmood

doi:10.4236/scd.2016.61001

Abstract

In the brain, there are hundreds of types of specialized neurons and to generate one type of them we need to have neural progenitors for differentiation to specific neuron type. Mesenchymal stem cells (MSCs) are easily isolated, cultured, manipulated ex vivo, showing great potential for therapeutic applications. The adult MSCs have the potential to produce progeny that differentiate into a variety of cell types such as neurons. This fact suggests that MSCs derived neurons are an important cell type and a deep understanding of the molecular characteristics of it would significantly enhance the advancement of cell therapy for neurological disorders. Therefore, in this study, we isolated, identified, and studied neural progenitors by measuring expression levels through neurogenesis pathway of three neural differentiation markers nestin (NES), neurofilament (NF-L), and microtubule association protein (MAP-2) from mouse bone marrow MSCs (mouse bmMSCs) by using butylated hydroxyanisole (BHA) and diethyl sulfoxide (DMSO) as neural inducers agents. The results of immunocytochemistry and Real Time-PCR showed that in contrast to MSCs, neural differentiated cells showed neural progenitor pattern by showing stable increase in NES gene expression through differentiation process with increasing the protein expression through different exposures times, while NF-L gene and protein expression start to increased after 48 h but not replaced the NES expression completely even when its expression passed NES levels. The maturation marker Map-2 expression was low during the duration of differentiation period in protein and gene expression, which prove that these cells are still progenitors and can be redirected into specific type of neurons by further treatments.

Highlights

Mesenchymal stromal cells or Mesenchymal stem cells (MSCs) are a type of widely distributed adult stem cell in connective tissues [1] [2]
After 24 h, only a few cells attached to the plastic culture flasks sparsely, and formed adherent cells while the non-adherent cells were discarded by the first medium change usually after 24 or 48 h (Figure 1(A))
Whereas the MAP-2 proteins showed the less expression levels compared with NES and NF-L with significant difference between different time at level at p < 0.05 compared with control. These results indicated the activity of butylated hydroxyanisole (BHA) in the neurogenesis stages of MSCs toward the neural differentiation cells

Summary

Introduction

Mesenchymal stromal cells or MSCs are a type of widely distributed adult stem cell in connective tissues [1] [2] They were easy to isolate and propagate and possess the ability to differentiate into many different cell lineages, including osteoblasts, chondrocytes, adipocytes, hepatocytes, and neuron-like cells since the nineteenth of the past century [3]-[6]. With these properties, MSCs have been widely applied in regenerative medicine research and experimental cell therapy for a wide range of disorders such parkinson’s disease (PD) [7], huntington’s disease [8], amyotrophic lateral sclerosis (ALS) [9], and alzheimer’s disease (AD) [10]. Protein expression levels of nestin, β III-tubulin, and tyrosine hydroxylase remarkably increased in differentiated MSCs [22]

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