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Preliminary evidence for the dedifferentiation of RAW 264 cells into mesenchymal progenitor-like cells by a purine analog.
MedLine Citation:
PMID:  22519969     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Dedifferentiation of cells to multipotential cells is of interest since they have potential regenerative capacity. Our purpose was to de- and re-differentiate murine RAW 264.7 cells, a committed macrophage cell line of hematopoietic origin, into mesenchymal-like cells such as osteoblasts. RAW 264.7 cells in culture were treated with 5µM reversine, a purine analog that was shown to dedifferentiate myoblasts in osteoblasts. Treatment with reversine resulted in a significant increase in the expression of the STRO-1 antigen, a marker of mesenchymal stem/progenitor cells: from 0.6 ± 0.5 % cells in untreated RAW cells to 19.0 ± 8.6 % in treated cells, but there was no increase in the expression of SH-2 (CD105), an earlier marker of mesenchymal stem cells. The effects of reversine were significantly curtailed by 67% when cultures were pre-treated with the JNK pathway blocker SP600125. These STRO-1+ cells retained a multipotential status and were capable of re-differentiating into cells with osteogenic and lipogenic characteristics under inductive conditions. We showed that STRO-1+ cells in osteogenic medium significantly increased expression of the osteoblast marker osteocalcin, and formed mineralized nodules. When seeded on a demineralized scaffold of human bone in vitro, these cells deposited a calcium matrix. Under adipogenic conditions, expression of the adipocyte marker PPARγ2 on STRO-1+ cells was elevated and cultures stained positive with oil red O. Our results demonstrated that treating a committed hematopoietic cell line with a purine analog can alter cell development and result in cellular reverse-transformation into stage-limited multipotential cells. These cells could subsequently be re-differentiated into cells with characteristics of the mesenchymal-lineage, such as those of an osteoblast and/or adipocyte, under inductive conditions.
Authors:
Guowei Qu; Herb P von Schroeder
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-4-20
Journal Detail:
Title:  Tissue engineering. Part A     Volume:  -     ISSN:  1937-335X     ISO Abbreviation:  -     Publication Date:  2012 Apr 
Date Detail:
Created Date:  2012-4-23     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101466659     Medline TA:  Tissue Eng Part A     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
University of Toronto, Faculty of Dentistry, Toronto, Ontario, Canada; guoweiqu@hotmail.com.
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