Gene Expression Profile during Chondrogenesis in Human Bone Marrow derived Mesenchymal Stem Cells using a cDNA Microarray

Abstract

Mesenchymal stem cells (MSCs) have the capacity to proliferate and differentiate into multiple connective tissue lineages, which include cartilage, bone, and fat. Cartilage differentiation and chondrocyte maturation are required for normal skeletal development, but the intracellular pathways regulating this process remain largely unclear. This study was designed to identify novel genes that might help clarify the molecular mechanisms of chondrogenesis. Chondrogenesis was induced by culturing human bone marrow (BM) derived MSCs in micromass pellets in the presence of defined medium for 3, 7, 14 or 21 days. Several genes regulated during chondrogenesis were then identified by reverse transcriptase-polymerase chain reaction (RT-PCR). Using an ABI microarray system, we determined the differential gene expression profiles of differentiated chondrocytes and BM-MSCs. Normalization of this data resulted in the identification of 1,486 differentially expressed genes. To verify gene expression profiles determined by microarray analysis, the expression levels of 10 genes with high fold changes were confirmed by RT-PCR. Gene expression patterns of 9 genes (Hrad6B, annexinA2, BMP-7, contactin-1, peroxiredoxin-1, heat shock transcription factor-2, synaptotagmin IV, serotonin receptor-7, Axl) in RT-PCR were similar to the microarray gene expression patterns. These findings provide novel information concerning genes involved in the chondrogenesis of human BM-MSCs.

Identification of new genes associated to senescent and tumorigenic phenotypes in mesenchymal stem cells

“…Transcriptome studies of hMSCs have focused on differential expression patterns among cells obtained from different sources 19 – 26 , different stages of the differentiation process 27 – 30 , and different cultivation times 31 – 35 . Differentially expressed genes have already been identified in bone marrow stem cells (hBMSC) at the 20 th passage compared to the 1st passage, adipose tissue stem cells (ASCs) at the 30th passage compared to the 1st passage 31 , hBM-MSC at the 15th passage compared to the 7th passage 32 , umbilical cord mesenchymal stem cells (UC-MSC) at the 15th passage compared to the 3rd passage 33 , hBMSCs at 33 population doubling levels (PDL) compared to 3 PDL 34 , and in BMSC at the 15th passage compared to 10th passage 35 .…”

Human chondrocyte migration behaviour to guide the development of engineered cartilage

“…Current methods for human cells reported in the literature employ various passage conditions and culture media formulae to support growth of chondrocytes (e.g. (Adesida et al , ; Alves da Silva et al , ; Bian et al , ; de Mara et al , ; Ghone and Grayson, ; Handorf and Li, ; Matsumoto et al , ; Mendelson et al , ; Park et al , ; Perrier et al , ; Wu et al , ; Yin et al , ; Yoo et al , )).…”

Bone growth in juvenile rhesus monkeys is influenced by 5HTTLPR polymorphisms and interactions between 5HTTLPR polymorphisms and fluoxetine

“…In addition, in a reanalysis of the data which included 5HTTLPR polymorphism genotyping, an interaction between 5HTTLPR polymorphisms and SSRI treatment was found for the total lumbar BMD z-score [24]. Effects of SSRIs on chondrocytes and cartilaginous tissue growth have not been studied, but serotonin receptors have been identified in embryonic and adult cartilaginous tissue [55, 56]. …”