Team:NYMU-Taipei/ymivenusianrd.html

Result & Discussion

There were three different types of cells we chose to do the coculture experiment. Part 1 revealed that J774 macrophage cell line enabled the entry of cyanobacteria by phagocytosis. Part2 and part3 were both mouse induced pluripotent stem cells but in different stage of stem cells. Secondly was non-EB iPS cells, separated from mouse embryonic fibroblasts (MEFs) and subcultured on culture dish. Thirdly was iPS cells in the Embryoid body (EB) stage. Later experiments showed three parts of results, J774 macrophage cell line(figure 2), non-EB iPS cells(figure 3), and iPS cells at the EB stage(figure 4). Red pots were S. elongates PCC7942 under fluorescence microscope 530-550 nm excitation filter. Under the transmitted light, transparent one was cells.

Part 1

Figure 2 revealed coculturing results of J774 macrophage cell lines and S. elongatus PCC 7942(wildtype/inv-llo). After 24-hour coculturing and 24-hour cell culture under gentamycin 100 ug/ml, same concentration of wildtype and inv-llo transformed type(both were OD 0.05) both entered into J774 macrophage cell line by phagocytosis and the help of inv-llo.

In figure 2,(B)(D) were the images under the fluorescence microscope 530-550 nm excitation filter. Red pots were S. elongates PCC7942. Under the transmitted light, round and transparent one was J774 macrophage cell line. (A)(C) were the merged images of bright fields and fluorescence. Compared inv-llo transformed type to wildtype, red pots of inv-llo transformed type were less bright than those of wildtype. Additionally, inv-llo transformed type numbers in cells were less than wildtype numbers. It might suggest that inv-llo gene could raise the entry speed and the escape rate.

Part 2

Figure 3 showed the coculture results of non-EB iPS cells. Cells and S.elongatus PCC7942(wildtype/inv-llo). Images(A)(B)(E)(F)(G)(H) were inv-llo strain and non-EB iPS cells coculturing results (A,B: under fluorescence microscope E~H: under confocal laser microscope.) Cells and cyanobacteria at the separate focus revealed the negative symbiosis. However, inv-llo plasmid showed that inv-llo would recruit cyanobacteria to cells. 3D image under the confocal laser microscope(image H) revealed the recruitment of cyanobacteria.

Part 3

Figure 4 revealed the coculture results of S.elongatus PCC 7942 (wt/inv-llo strain)and iPS cells at EB stage.

There were similar and relative results of the iPS cells at EB stage and non-EB iPS cells. Both inv-llo type of S.elongatus PCC7942 didn’t enter into stem cells but triggered the recruitment to cells. Because embryoid body contained not only one cell, the entry for cyanobacteria would be more difficult but the recruitment provided a potential for invading mammalian cells. Future study will focus on different coculturing condition and after invasion into stem cells, differentiation could render Venusian.