The characteristic of mesenchymal stem cells from peripheral blood and mobilized peripheral blood in comparison to bone marrow derived mesenchymal stem cells
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Date
2010-03-10
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Thammasat University, Rangsit Campus
Abstract
Objective: To isolate and characterize mesenchymal stem cells (MSCs) from peripheral blood and G-CSF mobilized peripheral blood. Methods: Mononuclear cells (MNCs) were isolated from peripheral blood, G-CSF mobilized peripheral blood and bone marrow using gradient centrifugation. The numbers of MSCs in these three sources were quantified using flow cytometry. The isolated MNCs were then cultured to generate MSCs. The MSCs generated from those three sources were studied in term of the MSC marker (CD73, CD90, CD105 and CD106) expression, the ability to generate colony (CFU-F) in culture and the ability to differentiate toward osteocyte and adipocyte-lineages. Results: The percentage of cells that expressed CD90 in fresh MNC populations isolated from bone marrow (BM-MNCs), peripheral blood (PB-MNCs) and mobilized peripheral blood (MPB-MNCs) is 1.94, 2.1 and 0.05 respectively, while the expression of CD73 in BM-MNCs, PB-MNCs and MPB-MNCs is 5.2, 15.2 and 7.8 respectively. The percentage of cells that expressed CD105 in BM-MNCs, PB-MNCs and MPB-MNCs is 5.3, 4.1 and 2.75, respectively while the expression of CD106 in those three populations is 2.82, 2.36 and 4.5 respectively. The ability of BM-MNCs, PB-MNCs and MPB-MNCs to generate colony in culture (CFU-F) is 67, 30 and 48 colonies per 10⁶ plating MNCs, respectively. After culture for three passages, more than 64% of BM-MNCs, PB-MNCs and MPB-MNCs homogeneously expressed CD73, CD90 and CD105. In contrast, the expression of CD45 (marker of hematopoietic cells) in those populations is negative. In addition, the bone marrow-derived MSCs also have an ability to differentiate toward osteocyte and adipocyte-lineages. Conclusion: We have successfully isolated and characterized MSCs from both peripheral blood and G-CSF mobilized peripheral blood. Those MSCs expressed several MSC markers, including CD73, CD90, CD105 and CD106, and able to generate colonies in culture in a manner similar to those of BM-MSCs. Our results suggest that these PB-MSCs and MPB-MSCs might be used as an alternative source for the clinical treatment in the future.
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Thammasat Medical Journal - ธรรมศาสตร์เวชสาร; Vol.8 No.3 July-September 2008; 281-291