Difference between revisions of "Team:TecCEM/Results"

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     <img src="https://static.igem.org/mediawiki/2018/a/aa/T--TecCEM--Cells.gif" alt="Cell Gif">
 
     <h1>Results</h1>
 
     <h1>Results</h1>
 
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             <h1>Results</h1>
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             <h3>Project/ Results</h3>
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            <h4>Overview </h4>
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            <p>To accomplish our project objective several recombinant proteins must be produced in order to analyze their effects on a co culture of fibroblasts (L929) and mesenchymal cells  after an in vitro burn assay. To evaluate the efficiency of the treatment, a measurement of the proliferation rate was performed using LDH (lactate dehydrogenase) as an analysis metabolite. Given that our treatment involves the usage of a growth factor, the rate at which this protein is released into the medium is critical to avoid adverse effects in the cellular line (such as cancer), for that reason a nano encapsulation with chitosan was also performed to control the rate at which our growth factor is released.</p>
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        <a href="#experimentsSubmenu" data-toggle="collapse" aria-expanded="false" data-offset="100" data-change="sidemenu">
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            <span data-change="el" class="d-inline-block open"></span>
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            Index
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        </a>
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        <ul class="collapse list-unstyled" id="experimentsSubmenu">
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            <h3>Summary</h3>
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                <a data-target="#overview">Overview</a>
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            <li>
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                <a data-target="#summary">Summary</a>
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                            <img src="https://static.igem.org/mediawiki/2018/5/51/T--TecCEM--Gelprueba1.jpg" class="figure-img img-fluid rounded"
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                                alt="BOB-5">
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                            <figcaption class="figure-caption"><strong>Figure 4. Procedure for PDMS membrane production.</strong></figcaption>
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    <div class="content">
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        <div class="container-fluid first-container" id="description">
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            <div class="row" id="overview">
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                <div class="col">
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                    <h2>Overview </h2>
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                    <p>To accomplish our project objective several recombinant proteins must be produced in order to
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                        analyze
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                        their effects on a co culture of fibroblasts (L929) and mesenchymal cells after an in vitro
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                        burn assay.
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                        To evaluate the efficiency of the treatment, a measurement of the proliferation rate was
 +
                        performed
 +
                        using LDH (lactate dehydrogenase) as an analysis metabolite. Given that our treatment involves
 +
                        the
 +
                        usage of a growth factor, the rate at which this protein is released into the medium is
 +
                        critical to
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                        avoid adverse effects in the cellular line (such as cancer), for that reason a nano
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                        encapsulation with
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                        chitosan was also performed to control the rate at which our growth factor is released.</p>
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            <div class="row" id="summary">
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                    <h2>Summary</h2>
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                    <!-- <div class="text-center">
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                        <img src="https://static.igem.org/mediawiki/2018/5/51/T--TecCEM--Gelprueba1.jpg" class="figure-img img-fluid rounded"
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                            alt="BOB-5">
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                        <figcaption class="figure-caption"><strong>Figure 1. Procedure for PDMS membrane production.</strong></figcaption>
 
                         </figure>
 
                         </figure>
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                    <h3>What did we accomplish?</h3>
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                    <ul>
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                        <li>Nanoencapsulation in chitosan of leptin, BSA and RFP.</li>
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            <h3>What should this page contain?</h3>
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                        <li>Realization of in vitro burn assay.</li>
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            <ul>
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                        <li>Realization of leptin proliferation essay.</li>
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                <li> Clearly and objectively describe the results of your work.</li>
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                        <li>Obtaining parts with adequate enzyme recognition sites.</li>
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                <li> Future plans for the project. </li>
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                        <li>Protein production of leptin and tenascin C (More validations are needed).</li>
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                <li> Considerations for replicating the experiments. </li>
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                        <li>Co-culture of fibroblast and mesenchymal cells.</li>
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            </ul>
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                        <li>Cellular growth in TaCO-BOB hardware.</li>
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                    </ul>
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                    <h3>What happened?</h3>
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                    <ul>
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        <div class="col">
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                        <li>We started ligating parts, no bands of expected size were observed but we attributed this
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            <h3>Describe what your results mean </h3>
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                            fact to low sensitivity of agarose gels, so we proceeded with transformation and protein
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            <ul>
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                            induction. Nevertheless, we realized that consistent results were not achieved (bad protein
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                <li> Interpretation of the results obtained during your project. Don't just show a
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                            migration rates, sometimes no induction band could be observed). [Unfortunately, we
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                    plot/figure/graph/other,
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                            invested a lot of time on this period]</li>
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                    tell us what you think the data means. This is an important part of your project that the judges
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                        <li>Then we started thinking that our parts did not had the necessary extra nucleotides for
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                    will look
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                            proper cleavage of restriction enzymes and what we were truly observing was the effect of
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                    for. </li>
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                            star activity and inappropriate ligation, resulting in non specific plasmids which did not
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                <li> Show data, but remember all measurement and characterization data must be on part pages in the
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                            contained our whole part but conferred the bacteria antibiotic resistance. </li>
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                    Registry.
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                    </ul>
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                </li>
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                    <h3>How did we solved it?</h3>
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                <li> Consider including an analysis summary section to discuss what your results mean. Judges like to
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                    <p>We designed primers that incorporate the necessary nucleotides into our parts. Thus, we started
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                    read what
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                        having proper migrations of project parts and posterious transformations and protein production
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                    you think your data means, beyond all the data you have acquired during your project. </li>
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                        of BBLEP.</p>
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            </ul>
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                    <h3>How should we improve?</h3>
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                    <p>By including better protein reporters for the screening of transformed, functional bacteria
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                        colonies.</p>
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            <h3> Project Achievements </h3>
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            <p>You can also include a list of bullet points (and links) of the successes and failures you have had over
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                your
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                summer. It is a quick reference page for the judges to see what you achieved during your summer.</p>
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            <ul>
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                <li>A list of linked bullet points of the successful results during your project</li>
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                <li>A list of linked bullet points of the unsuccessful results during your project. This is about being
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                    scientifically honest. If you worked on an area for a long time with no success, tell us so we know
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                    where
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                    you put your effort.</li>
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            </ul>
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        <div class="col">
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            <h3>Inspiration</h3>
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            <p>See how other teams presented their results.</p>
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            <ul>
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                <li><a href="https://2014.igem.org/Team:TU_Darmstadt/Results/Pathway">2014 TU Darmstadt </a></li>
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                <li><a href="https://2014.igem.org/Team:Imperial/Results">2014 Imperial </a></li>
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                 <li><a href="https://2014.igem.org/Team:Paris_Bettencourt/Results">2014 Paris Bettencourt </a></li>
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             </ul>
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         </div>
 
         </div>
 
     </div>
 
     </div>

Revision as of 20:59, 17 October 2018

Cell Gif

Results

Overview

To accomplish our project objective several recombinant proteins must be produced in order to analyze their effects on a co culture of fibroblasts (L929) and mesenchymal cells after an in vitro burn assay. To evaluate the efficiency of the treatment, a measurement of the proliferation rate was performed using LDH (lactate dehydrogenase) as an analysis metabolite. Given that our treatment involves the usage of a growth factor, the rate at which this protein is released into the medium is critical to avoid adverse effects in the cellular line (such as cancer), for that reason a nano encapsulation with chitosan was also performed to control the rate at which our growth factor is released.

Summary

What did we accomplish?

  • Nanoencapsulation in chitosan of leptin, BSA and RFP.
  • Realization of in vitro burn assay.
  • Realization of leptin proliferation essay.
  • Obtaining parts with adequate enzyme recognition sites.
  • Protein production of leptin and tenascin C (More validations are needed).
  • Co-culture of fibroblast and mesenchymal cells.
  • Cellular growth in TaCO-BOB hardware.

What happened?

  • We started ligating parts, no bands of expected size were observed but we attributed this fact to low sensitivity of agarose gels, so we proceeded with transformation and protein induction. Nevertheless, we realized that consistent results were not achieved (bad protein migration rates, sometimes no induction band could be observed). [Unfortunately, we invested a lot of time on this period]
  • Then we started thinking that our parts did not had the necessary extra nucleotides for proper cleavage of restriction enzymes and what we were truly observing was the effect of star activity and inappropriate ligation, resulting in non specific plasmids which did not contained our whole part but conferred the bacteria antibiotic resistance.

How did we solved it?

We designed primers that incorporate the necessary nucleotides into our parts. Thus, we started having proper migrations of project parts and posterious transformations and protein production of BBLEP.

How should we improve?

By including better protein reporters for the screening of transformed, functional bacteria colonies.