Purpose
The aim of this study was to investigate the relation between stage-specific embryonic
antigen 4 (SSEA4) expression and synovium-derived stem cell (SDSC) lineage differentiation.
Methods
Human SDSCs were collected during arthroscopic surgery from 4 young patients with
anterior cruciate ligament injuries. Passage 2 SDSCs were sorted by fluorescence-activated
cell sorting using phycoerythrin-conjugated monoclonal antibody against SSEA4 into
3 groups: SSEA4(+) cells, SSEA4(−) cells, and unsorted control cells. After 1 more
passage, expanded cells from each group were evaluated for SSEA4 expression by use
of flow cytometry as well as multilineage differentiation capacities, including chondrogenesis,
adipogenesis, and osteogenesis, using biochemical analysis, histologic analysis, immunostaining,
and real-time polymerase chain reaction.
Results
After cell sorting, 1 more passage expansion decreased SSEA4(+) cells from 99.8% to
79.2% and increased SSEA4(−) cells from 4.4% to 53.3% compared with 70.3% in the unsorted
cell population. SSEA4(−) SDSCs with a lower cell proliferation exhibited higher chondrogenic
potential (in terms of the ratio of glycosaminoglycan to DNA [P < .001] and COL2A1 [type II collagen] messenger RNA [mRNA] [P < .001]) and adipogenic potential (in terms of oil red O staining and quantitative
assay [P = .007], LPL [lipoprotein lipase] mRNA [P = .005], and CEBP [CCAAT/enhancer-binding protein alpha] mRNA [P = .010]). In contrast, SSEA4(+) SDSCs retained cell expansion and enhanced osteogenic
capacity, as evidenced by intense calcium deposition stained by alizarin red S and
a significantly elevated expression of OPN (osteopontin) mRNA (P = .007).
Conclusions
In this study, for the first time, we showed the benefit of using the surface marker
SSEA4 in SDSCs to preferentially sort a mixed population of cells. SSEA4(+) SDSCs
indicated a strong potential for osteogenesis rather than chondrogenesis and adipogenesis.
Clinical Relevance
SDSC-based mesenchymal tissue regeneration can be easily achieved by arthroscopic
harvesting followed by quick cell sorting.
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Article info
Publication history
Accepted:
December 16,
2013
Received:
July 3,
2013
Footnotes
This project was partially supported by research grants from the West Virginia University Senate Research Grant Award (R-12-010), the AO Foundation (S-12-19P), the Musculoskeletal Transplant Foundation, and the National Institutes of Health R03 Program (1 R03 AR062763-01A1 and 5 R03 DE021433-02). The authors report that they have no conflicts of interest in the authorship and publication of this article.
Identification
Copyright
© 2014 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.
