Maintenance of “stem cell” features of cartilage cell sub-populations during in vitro propagation

Journal of Translational Medicine

Wilhelm K Aicher²  Juergen A Mollenhauer¹  Christian Freudigmann³  Claudia Stippich³  Karin Benz³ 

[1] Rush University Medical Center, Chicago, IL, USA;KFO273, Department of Urology, University of Tuebingen Hospital, Tuebingen, Germany;NMI Natural and Medical Sciences Institute at the University of Tuebingen, Markwiesenstr. 55, Reutlingen, 72770, Germany

关键词: Differentiation;    Magnetic sorting;    Autologous chondrocyte implantation;    Disc cells;    Chondrocytes;    Stem cells;   

Others  :  828141
DOI  :  10.1186/1479-5876-11-27

 received in 2012-09-14, accepted in 2013-01-21,  发布年份 2013

【 摘 要 】

Background

The discovery of mesenchymal stem cells (MSCs) or MSC-like cells in cartilage tissue does not tie in well with the established view that MSCs derive from a perivascular niche. The presence of MSCs may raise concerns about specificity and application safety, particularly in terms of the regulatory site. The aim of the present study was to investigate the benefits or possible risks of the MSC-like properties of cells isolated from cartilage in the context of autologous chondrocyte implantation.

Methods

Chondrocytic cells were isolated from cartilage or intervertebral disc tissue. Flow cytometry was used to analyze the expression of cell surface antigens. MSC-like cells were either enriched or depleted by means of magnetic cell sorting (MACS) involving the monoclonal antibodies W5C5/SUSD2 and W8B2/MSCA-1. We addressed the issues of prolonged expansion of such cells as well as the influence of culture medium as a trigger for selecting a single cell type. Established protocols were used to study in vitro differentiation. In addition to histological and biochemical assessment, the acquired phenotypes were also evaluated on the mRNA transcript level.

Results

In the studied cells, we found strongly analogous expression of antigens typically expressed on MSCs, including CD49e, CD73, CD90, CD105, CD140b and CD166. The expression of W5C5 and W8B2 antigens in cartilage cell sub-populations did not correlate with multi-potency. We demonstrated that a chondroid precursor, but not a bona fide multipotent mesenchymal, cell type can be obtained under established in vitro culture conditions. The culture media used for expansion influenced the cell phenotype.

Conclusions

The risk of adverse adipose or osseous differentiation is not posed by expanded chondrocyte cultures, even after enrichment of putative MSC-like cell populations by MACS. It is possible that this limited “stemness” in chondrocytes, expanded for use in ACI, may instead be beneficial as it allows re-differentiation under appropriate conditions despite prolonged times in culture.

【 授权许可】

   
2013 Benz et al.; licensee BioMed Central Ltd.

【 图 表 】

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