Screening and cloning the genes differentially expressed in human embryonic AGM-derived stromal cells

• Published in: Zhongguo shi yan xue ye xue za zhi Vol. 14; no. 4; p. 726
• Main Authors: Fu, Jin-Rong, Chen, Hui-Qin, Zhang, Xu-Chao, Huang, Shao-Liang, Zhou, Dun-Hua, Huang, Ke
• Format: Journal Article
• Language: Chinese
• Published: China 01.08.2006
• Subjects: Aorta - embryology; Cloning, Molecular; Embryonic Stem Cells - cytology; Embryonic Stem Cells - metabolism; Endothelial Cells - cytology; Endothelial Cells - metabolism; Gene Expression; Gene Expression Profiling; Gonads - embryology; Hematopoiesis - physiology; Hematopoietic Stem Cells - cytology; Humans; Mesonephros - cytology; Oligonucleotide Array Sequence Analysis; Stromal Cells - cytology; Stromal Cells - metabolism
• ISSN: 1009-2137

To screen and separate the genes differentially expressed in human embryonic aorta-gonad-mesonephros (AGM)-derived stromal cells, a subtracted library was generated through the suppression subtractive hybridization using the cDNA of human embryonic AGM-derived stromal cells as target and human fetal liver (FL)-derived stromal cells as drivers. Then a high though screening technique, gene chip, was used to screen the differentially expressed genes in the established subtractive library. Approximately 18 of the resulting subtracted cDNA clones were partially sequenced and analyzed by blastn in the GenBank database. The results showed that 211 Clones were selected and identified from the established subtractive library, the positive ratio was amount to 76.4%. 18 over-expressed genes were screened by gene chip with more than a 5-fold difference expression levels between AGM and FL-derived stromal cells, and were selected to sequence, results of sequencing indicated that the 18 sequences was compared to known sequ