Gene expression profile of the differentiation of bone marrow stromal stem cells into neuron-like cells before and after induction*,⋆

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Abstract

BACKGROUND:

Accumulating evidences demonstrate that bone marrow stromal stem cells (BMSCs) can in vitro differentiate into neuron-like cells, which may be good candidates transplanted for some neurodegenerative diseases. However, the involved molecular mechanism remains unclear.

OBJECTIVE:

To detect the valuable changes of the gene expression profile in the induced differentiation of BMSCs into neuron-like cells.

DESIGN, TIME AND SETTING:

This observational experiment was performed at the Brain Lab in the Second Affiliated Hospital of Soochow University from December 2006 to December 2007.

MATERIALS:

Inbred Wistar rats weighing 300 g approximately were used in this study.

METHODS:

In vitro cultured and purified rat BMSCs were induced to differentiate into neuron-like cells with basic fibroblast growth factor and epidermal growth factor. Before and 7 days after induction, RNA was extracted respectively. Affymetrix microarrays were applied to identify the differential gene expression profile. Subsequently, some key genes were selected and checked by real-time PCR.

MAIN OUTCOME MEASURES:

The following parameters were measured: Culture and induced differentiation of BMSCs; identification of neuron-like cells after the induction; detection of microarrays before and after induction; analysis of the microarray results; validation by real-time PCR

RESULTS:

Morphologically, BMSCs were successfully induced to become neuron-like cells, (69.04±5.63)% of which were positive for β-TubulinIII. The microarrays detected 215 genes were different expressed (125 up-regulated and 90 down-regulated). In this group, 13 genes in the literature reported to be closely associated with neural development were specially noticed, among which 7 were up-regulated and 6 were down-regulated. Real-time PCR showed that the expression patterns of four selected key genes (Bmp4, Robo2, Numb and Enc1) were in accordance with those disclosed by the microarray.

CONCLUSION:

In the induced differentiation procedure of BMSCs into neuron-like cells, some important genes related to neural differentiation were expressed differentially. Further study on these interesting genes may establish molecular foundation for genetically engineering of BMSCs.

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