Copyright
©The Author(s) 2015.
World J Stem Cells. Sep 26, 2015; 7(8): 1137-1144
Published online Sep 26, 2015. doi: 10.4252/wjsc.v7.i8.1137
Published online Sep 26, 2015. doi: 10.4252/wjsc.v7.i8.1137
Figure 1 Study design.
Bone marrow samples from 7 healthy volunteers were aliquoted and treated by different protocols. The cells were plated for passage culture or CFU-F assay. CFU-F: Colony-forming unit-fibroblast.
Figure 2 Colony-forming unit-fibroblast and mesenchymal stromal cell numbers of different groups.
A: CFU-F numbers per 106 nucleated cells from seven bone marrow samples; B-D: Bone marrow samples were diluted and split into 12 aliquots containing 107 nucleated cells. The attached cells at passage 0 (B: × 104), passage 1 (C: × 105) and passage 2 (D: × 105) were counted and summed up. The average and standard errors are shown. CFU-F: Colony-forming unit-fibroblast.
Figure 3 Representative morphological features of the colonies and the attached cells.
Bone marrow samples were treated by different protocols and the cells were maintained for 12 d. Colonies of fibroblast-like cells were stained with Wright-Gimsa solution and observed under a microscope with a lower (A) and a higher magnification field (B). CTR: Un-coagulated control; UK: Urokinase-treated clots; CUT: Mechanically-cut clots; CLOT: Untreated clots.
Figure 4 Surface marker analysis of the attached cells by flow cytometry.
The attached cells at passage 2 were collected for flow cytometric analysis. The antigenic markers are indicated. UK: Urokinase-treated clots; CUT: Mechanically-cut clots; CLOT: Untreated clots. The results are representative of 7 individual experiments.
Figure 5 Identification of in vitro osteogenesis and adipogenesis.
The attached cells at passage 2 were exposed to specific inductive conditions and stained with oil-red (upper panel) or ALP solutions. The results are representative of 7 individual experiments. ALP: Alkaline phosphatase. CTR: Un-coagulated control; UK: Urokinase-treated clots; CUT: Mechanically-cut clots; CLOT: Untreated clots.
- Citation: Wang HX, Li ZY, Guo ZK, Guo ZK. Easily-handled method to isolate mesenchymal stem cells from coagulated human bone marrow samples. World J Stem Cells 2015; 7(8): 1137-1144
- URL: https://www.wjgnet.com/1948-0210/full/v7/i8/1137.htm
- DOI: https://dx.doi.org/10.4252/wjsc.v7.i8.1137