Chondrogenic potential of mesenchymal stem cells from horses using a magnetic 3D cell culture system

Figure 1 Flow cytometric analysis of the expression of cell surface markers CD90, CD44 and CD34 by mesenchymal stem cells. Representative dot plots and histograms of three sources: Synovial fluid (SF), bone marrow (BM) and adipose tissue (AT). SF, BM and AT. A: SF control; B: SF CD90+CD44+; C: SF CD34+; D: SF histograms control (black line), CD34+ (red line) and CD90+CD44+ (blue line); E: BM control; F: BM CD90+CD44+; G: BM CD34+; H: BM histograms control (black line), CD34+ (red line) and CD90+CD44+ (blue line); I: AT control; J: AT CD90+CD44+; K: AT CD34+; L: AT histograms control (black line), CD34+ (red line) and CD90+CD44+ (blue line).

Figure 2 Average and median of the total concentration of soluble proteins, aggrecan, and the ratio of the aggrecan to soluble proteins of the microspheroids from different sources of mesenchymal stem cells. A: Mean concentration (×), median (-), and standard deviation of the concentration of soluble proteins present in microspheroids extracted from samples of synovial fluid-derived mesenchymal stem cells (SF-MSCs), bone marrow-derived mesenchymal stem cells (BM-MSCs), and adipose tissue-derived mesenchymal stem cells (AT-MSCs). ¹ outlier; ^aP < 0.05 indicates the statistical difference in relation to BM-MSCs and SF-MSCs; B: Mean concentration (×), median (-), and standard deviation of the concentration of aggrecan from the microspheroids extracted from the samples of SF-MSCs, BM-MSCs, and AT-MSCs. ¹ outlier; ^aP < 0.05 indicates the statistical difference in relation to BM-MSCs and SF-MSCs; C: Mean concentration (×), median (-), and standard deviation of the ratio of the concentration of aggrecan to soluble proteins of the microspheroids extracted from the samples of SF-MSCs, BM-MSCs, and AT-MSCs. ¹ outlier; ^aP < 0.05 indicates the statistical difference in relation to BM-MSCs. SF-MSCs: Synovial fluid-derived mesenchymal stem cells; BM-MSCs: Bone marrow-derived mesenchymal stem cells; AT-MSCs: Adipose tissue-derived mesenchymal stem cells.

Figure 3 Representative images of 4',6-diamidino-2-phenylindole staining, and aggrecan and keratan sulfate expression in synovial fluid-mesenchymal stem cells using immunofluorescence. A: 4',6-diamidino-2-phenylindole (DAPI) 4 d; B: DAPI 7 d; C: DAPI 14 d; D: DAPI 21 d; E: Aggrecan 4 d; F: Aggrecan 7 d; G: Aggrecan 14 d; H: Aggrecan 21 d; I: Keratan sulfate 4 d; J: Keratan sulfate 7 d; K: Keratan sulfate 14 d; L: Keratan sulfate 21 d.

Figure 4 Representative images of 4',6-diamidino-2-phenylindole staining, and aggrecan and keratan sulfate expression in Bone marrow-mesenchymal stem cells using immunofluorescence. A: 4',6-diamidino-2-phenylindole (DAPI) 4 d; B: DAPI 7 d; C: DAPI 14 d; D: DAPI 21 d; E: Aggrecan 4 d; F: Aggrecan 7 d; G: Aggrecan 14 d; H: Aggrecan 21 d; I: Keratan sulfate 4 d; J: Keratan sulfate 7 d; K: Keratan sulfate 14 d; L: Keratan sulfate 21 d.

Figure 5 Representative images of 4',6-diamidino-2-phenylindole staining, and aggrecan and keratan sulfate expression in adipose tissue - mesenchymal stem cells using immunofluorescence. A: 4',6-diamidino-2-phenylindole (DAPI) 4 d; B: DAPI 7 d; C: DAPI 14 d; D: DAPI 21 d; E: Aggrecan 4 d; F: Aggrecan 7 d; G: Aggrecan 14 d; H: Aggrecan 21 d; I: Keratan sulfate 4 d; J: Keratan sulfate 7 d; K: Keratan sulfate 14 d; L: Keratan sulfate 21 d.