Abstract
BackgroundIdentification of surface markers for prospective isolation of functionally homogenous populations of human skeletal (stromal, mesenchymal) stem cells (hMSCs) is highly relevant for cell therapy protocols. Thus, we examined the possible use of CD146 to subtype a heterogeneous hMSC population.MethodsUsing flow cytometry and cell sorting, we isolated two distinct hMSC-CD146+ and hMSC-CD146− cell populations from the telomerized human bone marrow-derived stromal cell line (hMSC-TERT). Cells were examined for differences in their size, shape and texture by using high-content analysis and additionally for their ability to differentiate toward osteogenesis in vitro and form bone in vivo, and their migrational ability in vivo and in vitro was investigated.ResultsIn vitro, the two cell populations exhibited similar growth rate and differentiation capacity to osteoblasts and adipocytes on the basis of gene expression and protein production of lineage-specific markers. In vivo, hMSC-CD146+ and hMSC-CD146− cells formed bone and bone marrow organ when implanted subcutaneously in immune-deficient mice. Bone was enriched in hMSC-CD146− cells (12.6 % versus 8.1 %) and bone marrow elements enriched in implants containing hMSC-CD146+ cells (0.5 % versus 0.05 %). hMSC-CD146+ cells exhibited greater chemotactic attraction in a transwell migration assay and, when injected intravenously into immune-deficient mice following closed femoral fracture, exhibited wider tissue distribution and significantly increased migration ability as demonstrated by bioluminescence imaging.ConclusionOur studies demonstrate that CD146 defines a subpopulation of hMSCs capable of bone formation and in vivo trans-endothelial migration and thus represents a population of hMSCs suitable for use in clinical protocols of bone tissue regeneration.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-015-0266-z) contains supplementary material, which is available to authorized users.
Highlights
Identification of surface markers for prospective isolation of functionally homogenous populations of human skeletal stem cells is highly relevant for cell therapy protocols
We recently reported that canonical osteoblastic markers are not predictive for in vivo bone-forming capacity of hMSCs or hMSC “stemness” [8] and that there exist, in mesenchymal stem cell (MSC) cultures, cell populations committed to adipocyte or osteoblast lineages [9]
After sorting of the populations and expression of canonical mesenchymal stem cell (MSC) markers: CD44, CD63, CD73, CD105, CD14, CD34, and Cluster of differentiation 146 (CD146). c Analysis of population doublings demonstrating no significant difference between the different cell populations
Summary
Identification of surface markers for prospective isolation of functionally homogenous populations of human skeletal (stromal, mesenchymal) stem cells (hMSCs) is highly relevant for cell therapy protocols. A number of studies have isolated and characterized distinct populations of BM hMSCs by using a number of surface markers (e.g., Stro-1 and CD105 [13], CD271 [14], and CD56 [15, 16], and alkaline phosphatase (ALP) [17]). These markers enrich for an hMSC population with trilineage differentiation and colony-forming abilities, the isolated cells were still heterogeneous with respect to differentiation potential. Differences and similarities of the CD146− and CD146+ populations of hMSCs have not been totally clarified
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