Differentiation of Human Umbilical Cord Mesenchymal Stem Cells into Neural Stem Cell-like Cells Induced by Recombinant Human Peroxiredoxin-5

OBJECTIVE  To investigate the feasibility and possible mechanism of human umbilical cord mesenchymal stem cells(hUC-MSCs) differentiation into neural stem cell(NSC)-like cells induced by recombinant human peroxiredoxin-5(hPRDX5).

METHODS  Umbilical cords from normal full-term neonates were obtained under sterile conditions, and hUC-MSCs were cultured and identified. hUC-MSCs at passage 4 were divided into 5 groups: blank control group, 1 µg·mL⁻¹ hPRDX5, 10 µg·mL⁻¹ hPRDX5, positive control group (N2, B27, bFGF, and EGF), positive control+1 µg·mL⁻¹ hPRDX5, to investigate the potential of hPRDX5 in inducing hUC-MSCs differentiation into NSC-like cells. Cell morphology was observed using an inverted microscope. Immunohistochemistry was performed to detect the NSC marker Nestin. Immunofluorescence staining was used to detect microtubule-associated protein 2 (MAP2). Western blotting was employed to detect the expression of Nestin and toll-like receptor 4 (TLR4), a signaling pathway-related protein.

RESULTS  hUC-MSCs expressed high levels of CD29, CD73, CD90, and CD105, but did not express CD19, CD34, CD45, or HLA-DR. Following osteogenic, adipogenic, and chondrogenic differentiation, positive staining was observed with Alizarin red, oil red O, and Alcian blue, respectively. NSC-like cells induced by hPRDX5 showed positive immunostaining for Nestin. Furthermore, these hPRDX5-induced NSC-like cells differentiated into MAP2-positive neuron-like cells. The expression of TLR4, a signaling pathway-related protein, was significantly increased in NSC-like cells.

CONCLUSION  hPRDX5 can promote the differentiation of hUC-MSCs into NSC-like cells, and the underlying mechanism may be achieved through the TLR4 signaling pathway.