title: Analysis of mesenchymal cells derived from an chondrodysplasia punctuate patient and donors.
abstract: Conradi-Hunermann syndrome (CDPX2) is X-linked dominant disorder appeared with aberrant punctuate calcification. The skeletal cells derived from the marrow stroma are active in maintaining the skeletal formation. We obtained mesenchymal stem cells from a patient with CDPX2 and studied the formation of colony forming unit-fibroblast (CFU-F) in vitro in comparison cells obtained from normal donors. Cultured cells were studied morphologically and subjected to gene expression analysis. Marrow stromal cells (MSC)-chondrodysplasia punctuate (CDP) cells from CDPX2 were identified by their mosaic morphology formed three phenotypically distinct types of CFU-F colonies. One type consisted of normal fibroblasts with developed cell body and cellular processes; the second type contained pathological small cells without processes; and the third type comprised of mixed cells. We compared gene expression by the MSC-CDP to cells from normal donors. Transcription factors analyzed proliferation potential were similar in both normal and mixed colonies of MSC-CDP and similar to normal MSCs. The message expression for cytokines and extra cellular matrix (ECM) proteins revealed similar expression for biglycan, osteocalcin, and osteonectin, while IL-6, IL-11, and M-CSF mRNA levels were significantly higher in normal cells than in MSC-CDP. Mixed cells had elevated levels for IL-6 and M-CSF mRNA, but expressed IL-11 at the normal range. The studied genes were expressed at lower levels by the pathological (MSC-CDP) cells compared to normal ones. Hence, MSC-CDP was demonstrated to display abnormal morphology and transcription of several investigated genes. This study further illuminates the basis of the mosaic pattern of mesenchymal cells derived from a patient affected with CDPX2, and their gene expression involvement.
authors.affiliation: Department of Cell and Developmental Biology, Sackler School of Medicine, Tel-Aviv University, Tel-Aviv 66978, Israel.
authors.forename: Irena
authors.initials: I
authors.lastname: Shur
authors.forename: Orit
authors.initials: O
authors.lastname: Reish
authors.forename: Eli
authors.initials: E
authors.lastname: Ezra
authors.forename: Dafna
authors.initials: D
authors.lastname: Benayahu
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D001853:Bone Marrow / Q000378:metabolism; D002806:Chondrodysplasia Punctata / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D003114:Colony-Forming Units Assay; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D020869:Gene Expression Profiling; D006801:Humans; D007223:Infant; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D017154:Stromal Cells / Q000378:metabolism; D014019:Tissue Donors
chemical_list: D015415:Biomarkers