Team:NYMU-Taipei/ymivenusianrd.html

From 2012.igem.org

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       <td width="313" valign="top"><p><img width="298" height="224" src="http://igem.org/wiki/images/7/74/Ymivenusianrd_clip_image002.jpg" alt="描述: Tv9 [1]" /> <br />
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         (A)inv-llo    strain, merged image of fluorescence and bright field</p></td>
         (A)inv-llo    strain, merged image of fluorescence and bright field</p></td>
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       <td width="321" valign="top"><p><img width="306" height="230" src="http://igem.org/wiki/images/7/71/Ymivenusianrd_clip_image004.jpg" alt="描述: Tv9" /> <br />
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         (B)    inv-llo strain, image of fluorescence </p></td>
         (B)    inv-llo strain, image of fluorescence </p></td>
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       <td width="313" valign="top"><p><img width="298" height="224" src="http://igem.org/wiki/images/8/82/Ymivenusianrd_clip_image006.jpg" alt="描述: Tv2 [1]" /> <br />
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         (C)    wildtype, merged image of fluorescence and bright field</p></td>
         (C)    wildtype, merged image of fluorescence and bright field</p></td>
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       <td width="321" valign="top"><p><img width="296" height="222" src="http://igem.org/wiki/images/2/2c/Ymivenusianrd_clip_image008.jpg" alt="描述: Tv2" /> <br />
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         (D)    wildtype, image of fluorescence</p></td>
         (D)    wildtype, image of fluorescence</p></td>
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     <td width="318" valign="top"><p><img width="273" height="204" src="http://igem.org/wiki/images/7/75/Ymivenusianrd_clip_image002_0000.jpg" alt="描述: Tv5 [1]" /> <br />
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       (A)    inv-llo strain, merged image of fluorescence and bright field</p></td>
       (A)    inv-llo strain, merged image of fluorescence and bright field</p></td>
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     <td width="296" valign="top"><p><img width="279" height="209" src="http://igem.org/wiki/images/b/bb/Ymivenusianrd_clip_image004_0000.jpg" alt="描述: Tv5" /> <br />
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       (B)    inv-llo strain, image of fluorescence</p></td>
       (B)    inv-llo strain, image of fluorescence</p></td>
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     <td width="318" valign="top"><p><img width="273" height="204" src="http://igem.org/wiki/images/1/1e/Ymivenusianrd_clip_image006_0000.jpg" alt="描述: Tv1 [1]" /> <br />
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       (C)    wildtype, merged image of fluorescence and bright field</p></td>
       (C)    wildtype, merged image of fluorescence and bright field</p></td>
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       (D)    wildtype, image of fluorescence</p></td>
       (D)    wildtype, image of fluorescence</p></td>
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       <p>(E) inv-llo strain, merged image    of confocal microscope and bright field, 1000X</p></td>
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        (E) inv-llo strain, merged image    of confocal microscope and bright field, 1000X</p></td>
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      (F) inv-llo strain, image of confocal    microscope, 1000X </p></td>
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        (F) inv-llo strain, image of confocal    microscope, 1000X </p></td>
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     <td width="318" valign="top"><p><img src="http://igem.org/wiki/images/a/a0/Ymivenusianrd_clip_image016.jpg" alt="" width="271" height="271" /><br />
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       (G) inv-llo strain, image of    bright field, 1000X</p></td>
       (G) inv-llo strain, image of    bright field, 1000X</p></td>
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     <td width="296" valign="top"><p><img width="282" height="254" src="http://igem.org/wiki/images/0/04/Ymivenusianrd_clip_image018.jpg" alt="描述: ips3D調" /> <br />
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       (H)    inv-llo strain, 3D image under confocal microscope, 1000X</p></td>
       (H)    inv-llo strain, 3D image under confocal microscope, 1000X</p></td>
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       (A)    iPS cells at EB stage, image of bright field</p></td>
       (A)    iPS cells at EB stage, image of bright field</p></td>
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     <td width="312" valign="top"><p><img width="298" height="224" src="http://igem.org/wiki/images/4/45/Ymivenusianrd_clip_image004_0001.jpg" alt="描述: Tv4" /> <br />
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       (B)inv-llo    strain, image of fluorescence</p></td>
       (B)inv-llo    strain, image of fluorescence</p></td>
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     <td width="304" valign="top"><p><img width="290" height="217" src="http://igem.org/wiki/images/0/0e/Ymivenusianrd_clip_image006_0001.jpg" alt="描述: Tv1 [1]" /> <br />
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       (C)inv-llo    strain, merged image of fluorescence and bright field</p></td>
       (C)inv-llo    strain, merged image of fluorescence and bright field</p></td>
     <td width="312" valign="top"><p>Figure    4.<br />
     <td width="312" valign="top"><p>Figure    4.<br />
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<p>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&rsquo;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.</p>
<p>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&rsquo;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.</p>
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Revision as of 17:19, 17 October 2012

NYMU iGEM

Becoming Venusian !

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.


(A)inv-llo strain, merged image of fluorescence and bright field


(B) inv-llo strain, image of fluorescence


(C) wildtype, merged image of fluorescence and bright field


(D) wildtype, image of fluorescence

Figure 2. Red pots were S. elongates PCC7942.  Round and transparent one was J774 macrophage cell line. By phagocytosis, wildtype and inv-llo type entered into cells.

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.


(A) inv-llo strain, merged image of fluorescence and bright field


(B) inv-llo strain, image of fluorescence


(C) wildtype, merged image of fluorescence and bright field


(D) wildtype, image of fluorescence

描述: ips_C001Z021

 

 

 

 

 



(E) inv-llo strain, merged image of confocal microscope and bright field, 1000X













(F) inv-llo strain, image of confocal microscope, 1000X


(G) inv-llo strain, image of bright field, 1000X


(H) inv-llo strain, 3D image under confocal microscope, 1000X

Figure 3. Red pots were S. elongates PCC7942 under fluorescence microscope.  Transparent one was non-EB iPS cells. Both wildtype and inv-llo type could not enter into cells but inv-llo type could trigger itself recruitment to cells.

Part 3

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


(A) iPS cells at EB stage, image of bright field


(B)inv-llo strain, image of fluorescence


(C)inv-llo strain, merged image of fluorescence and bright field

Figure 4.
Red pots were S. elongates PCC7942 under fluorescence microscope. Gray one was iPS cells at the EB stage. Results showed that not many inv-llo PCC 7942 could enter into cells.

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.