Changes in the expression of CD106, osteogenic genes, and transcription factors involved in the osteogenic differentiation of human bone marrow mesenchymal stem cells

Abstract

Mesenchymal stem cells (MSCs) are well known to possess multipotential differentiation and are becoming a good tool for clinical research. However, specific markers for their purification and the mechanism of their osteogenic differentiation remain to be elucidated. In the present study, we compared the expression of CD106, and osteogenic differentiation-related proteins and genes in human bone marrow (BM)-derived MSCs, before and after differentiation by FACS, histochemical staining, immunohistochemical staining, RT-PCR, and real-time PCR. It was found that MSCs were positive for CD13, CD29, CD44, CD73, CD90, CD105, and CD166, but negative for CD14, CD31, CD34, CD62E, CD45, and GlyA. Notably, CD106 was detected before osteogenic induction, but its expression was downregulated 10 fold after 2 weeks of osteogenic differentiation as determined by flow cytometry. The results of RT-PCR and real-time PCR revealed that the expression of CD106 mRNA in MSCs significantly decreased by 7.1-, 4.2-, and 5.1-fold, respectively after osteogenic, chondrogenic, and adipogenic differentiation. In contrast, other MSC-positive markers described above did not change significantly even after differentiation. Compared to levels in control cells, after 2 weeks of osteogenic differentiation, mRNA levels of alkaline phosphatase, bone sialoprotein, osteocalcin, and transcript factors RUNX2 and Osterix showed more than 2-fold, 5-fold, 1.5-fold, 2-fold, and 5-fold increase, respectively. Thus, we speculate that CD106 might be a useful surface marker for BMMSCs. Moreover, alkaline phosphatase, type I collagen, osteonectin, osteopontin, and biglycin were expressed in the early stages of osteogenic differentiation before bone sialoprotein and osteocalcin. The present study should help to provide a novel marker for isolating purified MSCs and characterizing osteogenic differentiation.

Adipogenic, Chondrogenic and Osteogenic Differentiation of Clonally Derived Human Dermal Fibroblasts

“…The CD marker profile can be used in order to further characterize and purify the sccFB populations. The lack of CD106 expression in the present study suggests that the cells investigated are not mesenchymal stem cells according to Liu et al [2008].…”

“…The CD markers chosen have previously been employed to study mesenchymal stem cells [Pittenger et al, 1999;Zuk et al, 2002;Liu et al, 2008]. A number of CD markers included are specific for hematopoietic cells and were used in order to exclude any progenitor cells derived from circulating stem cell populations.…”

Resorbable electrospun polydioxanone fibres modify the behaviour of cells from both healthy and diseased human tendons

“…CD106 (VCAM-1) is a membrane protein of the immunoglobulin super family and can be unregulated in response to proinflammatory TNF-alpha and IL-1 (Simmons et al, 1992). Expression of CD106 gene was also shown to decrease after osteogenic differentiation of mesenchymal stem cells (Liu et al, 2008). We found that a greater proportion of diseased tendon cells were CD106 neg compared to hamstring cells.…”

Development of Novel Monoclonal Antibodies that Define Differentiation Stages of Human Stromal (Mesenchymal) Stem Cells

“…The cell surface glycoprotein CD44 is known to be expressed on MSCs and differentiated osteogenic descendants (Liu et al, 2008), and is often used in conjunction with other molecules to confirm MSC identity (Bianco et al, 2001). However, CD44 is a widely distributed molecule and not specific for MSC .…”