Basic Fibroblast Growth Factor and Epithelial Growth Factor could Induce Trans-differentiation of Mesenchymal Stem Cell into Neural Stem Cell

Abstract

Background & Aim Background Stem cell therapy has been considered as a potential strategy in regenerative for the patient with neurological deficits. Mesenchymal Stem/Stromal Cells (MSC) have been extensively investigated as a treatment in stroke. However, the MSC lost its potency for neural repair in aged patients due to the limited numbers of Neural Stem Cell (NSC). NSC is essential for neurogenesis of the injured brain. In vitro studies have shown that MSCs which were cultured in the laboratory has the capability to transform into different cells. Adding stimulants such as growth factors into the culture media could induce the trans-differentiation of MSC into NSC. However, to produce NSC in the clinical setting on a large scale is still a challenge. Objective In this study, we like to evaluate the use of epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), heregulin B27, and N2 the trans-differentiation mechanism of umbilical cord MSC (UC-MSC) into NSC. Methods, Results & Conclusion Method MSC was transferred into a 24-well plate at 2 × 104 cells per well. Induction of NSC is done using DMEM containing N2/B27 supplement, 20 ng/ml epidermal growth factor (EGF), and 10 ng/ml basic fibroblast growth factor (bFGF). Identification of success trans-differentiation is analyzed using NeuroFlour CDr3 antibody. Result and Discussion MSC transdifferentiates into NSC as an important source of cell in therapy for neurological diseases. UC-MSC was induced using N2 to helps the cell to commit to differentiation and its survival. B27 is used for the long term survival of neurons cells and helps the NSC to grow and proliferates. bFGF beliefs as pre-inductive molecules that enhance neuronal differentiation and self-renewal. Upon bFGF stimulation, NGFβR and NRP1 will upregulate and promotes differentiation. On the contrary, EGF induces differentiation at lower concentrations. EGF is used for inducing the expression of the neuron and glial marker. To confirm the successful transdifferentiating process, the expression of CDr3 in the cell culture is analyzed. CDr3 positive result as the membrane-permeable fluorescent probe was shown in Figure 1. NeuroFluor CDr3 binds to human fatty acid-binding protein 7 (FABP7) which use to confirm the neural induction from human stem cells. Conclusion UC-MSC could be cultured and differentiate into NSC in vitro when supplemented with N2/B27, bFGF, and EGF, which may produce NSC in the clinical setting on a large scale. Stem cell therapy has been considered as a potential strategy in regenerative for the patient with neurological deficits. Mesenchymal Stem/Stromal Cells (MSC) have been extensively investigated as a treatment in stroke. However, the MSC lost its potency for neural repair in aged patients due to the limited numbers of Neural Stem Cell (NSC). NSC is essential for neurogenesis of the injured brain. In vitro studies have shown that MSCs which were cultured in the laboratory has the capability to transform into different cells. Adding stimulants such as growth factors into the culture media could induce the trans-differentiation of MSC into NSC. However, to produce NSC in the clinical setting on a large scale is still a challenge. In this study, we like to evaluate the use of epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), heregulin B27, and N2 the trans-differentiation mechanism of umbilical cord MSC (UC-MSC) into NSC. MSC was transferred into a 24-well plate at 2 × 104 cells per well. Induction of NSC is done using DMEM containing N2/B27 supplement, 20 ng/ml epidermal growth factor (EGF), and 10 ng/ml basic fibroblast growth factor (bFGF). Identification of success trans-differentiation is analyzed using NeuroFlour CDr3 antibody.