Differential expression gene of rat bone marrow mesenchymal stem cells and chondrocytes screened by gene chip technique

Ya FENG; Lijie ZHAI; Zhiqiang WANG

ABSTRACT

BACKGROUND:

Presently used biological factors for inducing bone marrow mesenchymal stem cells (BMSCs) do not obtain mature chondrocytes. Cartilage tissue engineering using BMSCs as seeds does not collect tissue-engineered cartilage, which has value in the clinic. The obtained tissue is cartilage-like tissue.

OBJECTIVE:

To screen differentially expressed genes between rat BMSCs and chondrocytes by microarray. DESIGN, TIME AND

SETTING:

The cytology gene study was conducted at the Central Laboratory, First Affiliated Hospital, Dalian Medical University in July 2007. MATERIALS A total of 8 healthy Sprague Dawley rats aged 2 months were obtained from Animal Experimental Center, Dalian Medical University. 27K Rat Genome Array chip was supplied by Bo’ao, Beijing, China.

METHODS:

Rat BMSCs and chondrocytes in the aural region were sterilely isolated and cultured in vitro. Total RNA was extracted and purified using Trizol one-step method, and transformed into double chain cDNA probe following reverse transcription. Cy5-dCTP and Cy3-dCTP were used to label BMSCs and chondrocytes, which were hybridized and washed. The fluorescent signals were scanned by a scanner. The values were analyzed and calculated by GenePix Pro 4.0 software. MAIN OUTCOME

MEASURES:

Gene expression spectrum chip hybridization results.

RESULTS:

Among the differentially expressed genes (2 times difference), BMSCs as controls, upregulated and downregulated genes were 1 226 and 888, respectively. There were many differential expression gene of BMSCs and chondrocytes. Cy5/Cy3 20 genes are defined as significant differential expression gene. Thus, two important cytokines were found chondromodulin and connective tissue growth factor.

CONCLUSION:

The gene chip technique provides an ideal method for screening cytokines during study of tissue-engineered cartilage. Cartilage regulin and connective tissue growth factor highly express in chondrocytes, which indicated that the two have closely association with the differentiation of BMSCs into cartilage.