Isolation, expansion, and in vitro differentiation of human umbilical cord blood mesenchymal stromal cells into neural progenitor cells

Abstract:
Background And Objectives
Umbilical cord blood, as a source of mesenchymal stromal cells, has many advantages compared to other sources. This study aimed to provide a new method for the in vitro neural differentiation of human umbilical cord blood derived mesenchymal stromal cells (MSCs hUCB).
Materials And Methods
In this experimental study, MSCs hUCBs were isolated and characterized by morphologic, adipogenic, and osteogenic differentiation and immunophenotypical analysis. Then, neural induction of MSCs hUCB was performed by using RA, bFGF, NGF, EGF, AsA, IBMX, and neurobasal medium. Then, the relative expression of neural-specific genes was investigated by quantitative real-time PCR assays, REST 2009, and SPSS 11.5 software.
Results
Our results showed the osteocytic and adipocytic differentiation capacity and fibroblast-like morphology of MSCs hUCB. Flow cytometry analysis of MSCs hUCB revealed that the cells are positive for CD105 (78%), CD73 (78%), and negative for CD45 (2%), HLA-DR (2.5%). The cells showed the remarkable transition from fibroblast-like morphology to nueral progenitor cells. The results showed that the expression of GFAP, MBP, nestin, MAP-2 genes after neural induction significantly increased in comparison with that of the control as measured by quantitative real-time PCR assays (p < 0.05).
Conclusions
Treatment of MSCs hUCB with a set of growth factors and chemical materials induces neural differentiation and increases the efficiency of cell-based therapy for neurodegenerative diseases in the future. Although, the functionality of neural progenitor cells must be carefully assessed in animal models prior to use in clinical application.
Language:
Persian
Published:
Scientific Journal of Iranian Blood Transfusion Organization, Volume:12 Issue: 3, 2015
Pages:
266 to 276
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