Figure 1.
Spirulina protects against LPS induced decrease in cell proliferation.
(A) Results of the Stereological counts of the number of BrdU+ cells demonstrated that spirulina was able completely block the LPS induced decline in inflammation. *** p = 0.0049 (NIH-31 vs. NIH-31+LPS). ††† P = 0.0090 (NIH-31+LPS vs. spirulina+LPS). (B) Shows the representative BrdU staining in the rats without LPS. (C) After LPS in the rats fed the control diet there are fewer BrdU+ cells. (D) In the spirulina fed rat LPS did not cause a decrease in proliferation. (All photomicrographs taken at 10× magnification).
Figure 2.
BrdU found in microglia only in the control fed mice following LPS.
Low magnification confocal photomicrographs were used to determine if the BrdU was labeling non-neural cells. (A–C) In the control animals without LPS the majority of the BrdU cells were not found to be microglia. (D–E) In the control diet fed rats treated with LPS note that fewer BrdU cells were found and that many of the BrdU labeled cells were microglia. White arrow shown in high magnification in Fig. 3D–F Yellow arrow shown in high magnification in Fig. 3G–I. (G–I) In the spirulina fed rats the BrdU cells were not found to be microglia.
Figure 3.
Orthogonal projections of BrdU IBA1 co-staining.
(A–C) shows a representative cluster of BrdU+ cells in the SGZ confirming the cells are not microglia by the lack of colocalization with IBA-1. (D–E) shows a cluster of BrdU+ cells in the LPS treated rats that are not microglia. (G–I) shows a BrdU+ that does double label with the microglia marker IBA-1 as noted by the yellow color in the merged panel. (J–K) shows BrdU and IBA-1 staining in the Spirulina fed rats. Note, in this and all confocal images the side panels for each box are 90° rotations of the Z-stacks for confirmation of double labeling.
Figure 4.
Orthogonal projections of BrdU GFAP co-staining.
(A–D) Confocal images with orthogonal views demonstrate the absence of co-labeling of BrdU and GFAP in the 4 groups of animals examined. In the merged column you can observe co-localization of BrdU with DAPI as indicated by the purple. In constrast there is no colocalization of BrdU with GFP although the GFP labeled cells are closely associated with the BrdU positive cells. This supports our finding of no increased absolute numbers of GFAP labeled cells between any of the groups examined.
Figure 5.
Spirulina rats have a decreased astrocyte response to LPS.
(A) LPS caused a significant increase in the percentage of the dentate gyrus which was GFAP positive. The spirulina fed rats had a reduced GFAP response LPS compare to the control diet fed rats with LPS. *** p = 0.0002 (NIH-31 vs. NIH-31+LPS). ††† P = 0.0003 (NIH-31+LPS vs. spirulina+LPS). (B) There was no significant difference in the total number of GFAP+ cells. (C) Representative photomicrograph of the GFAP immunohistochemical staining in the hippocampus. The white arrow points to the area shown in (D) at 20× magnification.
Figure 6.
Spirulina increases proliferation of human hematopoetic stem cells in vitro.
(A) The effects of spirulina and NT-020 on the proliferation of human bone marrow cells was examined. Cells were plated in 96 well plates and spirulina at varying concentrations was added to the culture media. In a separate study shown in the same graph the highest dose of spirulina tested alone (125 ng/ml) was added to NT-020. After 72 hours in culture viability was tested using the MTT assay. Data is expressed at % over control, which are cells grown in media only. When spirulina is combined with NT-020 the effect is significantly higher than NT-020 alone was more than additive. (B) The effects of spirulina and NT-020 on the proliferation of human CD34+ cells was examined. Cells were plated in 96 well plates and spirulina at varying doses was added to the culture media, in a separate study shown in the same graph the highest dose of spirulina tested alone (125 ng/ml) was added to NT-020. After 72 hours in culture viability was tested using the MTT assay. Data is expressed at % over control, which is cells grown in media only. When spirulina is combined with NT-020 the effect is significantly higher than NT-020 alone (p<0.05 students 2-tailed t-test) and the effect appears to be additive. ** p<0.001 ***p<0.0001 (compare to control). P<0.0001 (NT-020 vs. spirulina+NT-020).
Figure 7.
Spirulina increases proliferation of human neural stem cells in vitro and protects against a TNFα insult.
Human neural progenitors grown under proliferation conditions were assessed by MTT assay (A) or BrdU (B) for the effects of spirulina (125 ng/ml) or NT-020 (500 ng/ml) or the two treatments combined in the presence or absence of TNFα (20 ng/ml) for 72 hours. (A) The MTT assay shows that spirulina alone or NT-020 alone increase proliferation; surprising, the in combination proliferation is decrease compare to control ** p<0.005(compare to media alone). Addition of 20 ng/ml human recombinant TNFα significantly decreased proliferation compare to media alone (***p<0.0001). Only the addition of spirulina alone was able to revert decrease in proliferation †††p<0.05 (compare to TNFα alone). (B) Spirulina and NT-020 alone significantly increased the number of BrdU+ cells. When added together spirulina and NT-020 cause a significant decrease in proliferation ** p<0.005 ***p<0.0001 (compare to media alone). The addition of TNFα significantly decreased proliferation compare to media alone (***p<0.0001). Spirulina alone was able to prevent the decrease in the number of BrdU+ cells. ††† p<0.0001 (compare to TNFα alone).