CD146/MCAM defines functionality of human bone marrow stromal stem cell populations

Harkness, Linda, Zaher, Walid, Ditzel, Nicholas, Isa, Adiba and Kassem, Moustapha (2016) CD146/MCAM defines functionality of human bone marrow stromal stem cell populations. Stem Cell Research and Therapy, 7 1: 4.1-4.13. doi:10.1186/s13287-015-0266-z


Author Harkness, Linda
Zaher, Walid
Ditzel, Nicholas
Isa, Adiba
Kassem, Moustapha
Title CD146/MCAM defines functionality of human bone marrow stromal stem cell populations
Journal name Stem Cell Research and Therapy   Check publisher's open access policy
ISSN 1757-6512
Publication date 2016-01-11
Year available 2016
Sub-type Article (original research)
DOI 10.1186/s13287-015-0266-z
Open Access Status DOI
Volume 7
Issue 1
Start page 4.1
End page 4.13
Total pages 13
Place of publication London, United Kingdom
Publisher BioMed Central
Language eng
Subject 2701 Medicine (miscellaneous)
1313 Molecular Medicine
1301 Biochemistry, Genetics and Molecular Biology (miscellaneous)
1307 Cell Biology
Abstract Background: 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. Thus, we examined the possible use of CD146 to subtype a heterogeneous hMSC population. Methods: Using 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. Results: In 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. Conclusion: Our 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.
Formatted abstract
Background: 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. Thus, we examined the possible use of CD146 to subtype a heterogeneous hMSC population.

Methods:
Using 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.

Results:
In 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, hMSCCD146+ 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.

Conclusion:
Our 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.
Keyword Bone marrow stem cells
CD146/MCAM
Osteogenic differentiation
Bone Formation
Q-Index Code C1
Q-Index Status Provisional Code
Grant ID 09-067112
2014-10309
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: HERDC Pre-Audit
Australian Institute for Bioengineering and Nanotechnology Publications
 
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Created: Wed, 13 Jan 2016, 02:43:02 EST by Linda Harkness on behalf of Aust Institute for Bioengineering & Nanotechnology