High-Yield Isolation, Expansion, and Differentiation of Rat Bone Marrow-Derived Mesenchymal Stem Cells with Fibrin Microbeads
Fibrin microbeads (FMB), made of extensively cross-linked dense and partially denatured fibrin, were used as a matrix for efficient isolation of mesenchymal stem cells (MSC) from rat bone marrow (BM). After 2 days of incubation of FMB with whole BM in suspension, a high number of cells of mesenchyma...
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Veröffentlicht in: | Tissue engineering 2006-08, Vol.12 (8), p.2343-2354 |
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Zusammenfassung: | Fibrin microbeads (FMB), made of extensively cross-linked dense and partially denatured fibrin, were
used as a matrix for efficient isolation of mesenchymal stem cells (MSC) from rat bone marrow (BM). After
2 days of incubation of FMB with whole BM in suspension, a high number of cells of mesenchymal origin
attached to the FMB. On the 14th day after their transfer to plastic, the yield of the cells isolated via FMB
was ∼3-4 times higher than that obtained by currently used protocols based solely on plastic adhesion.
This implies that the number of MSC in BM may be higher than previously reported. FACS analyses and
immunostaining showed the mesenchymal characteristics of these cells by positive staining for fibronectin,
vimentin, CD49E, and CD29. Immediately after isolation, less than 20% of the cells still expressed the
hematopoietic markers CD11b and CD45. Most of these cells were eventually eliminated after further
expansion of the isolated cells on plastic. Cells isolated via FMB were expanded in culture for more than
4 months and could be defined as MSC along this time period based on their ability to differentiate into
precursors of mesenchymal tissues, such as osteogenic, adipogenic, and chondrogenic cells. Similar differentiation
plasticity was observed in clones derived from single cells from whole MSC populations
isolated via FMB. Based on our results we propose that FMB can serve as a 3-dimensional biodegradable
matrix for isolation, differentiation, and possibly implantation of MSC for tissue regeneration. |
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ISSN: | 1076-3279 1557-8690 |
DOI: | 10.1089/ten.2006.12.2343 |