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<html xmlns="http://www.w3.org/1999/xhtml"> <head> <meta http-equiv="Content-Type" content="text/html; charset=utf-8" /> <title>NYMU iGEM</title> <link href="http://www.royals.com.tw/nymu/style.css" rel="stylesheet" type="text/css" /> <link href="http://www.royals.com.tw/nymu/css/navigation_js.css" rel="stylesheet" type="text/css" /> <script src="//ajax.googleapis.com/ajax/libs/jquery/1.8.1/jquery.min.js" type="text/javascript"></script> <script src="http://www.royals.com.tw/nymu/js/jquery.dimensions.js" type="text/javascript"></script> <script src="http://www.royals.com.tw/nymu/js/jquery.accordion.js" type="text/javascript"></script> <script type="text/javascript">  </script> </head> <body> <div class="ymi_spacer"></div> <div id="ymi_topcontainer"> <div id="ymi_top"> <a href="ymiproject.html" target="_parent">Project Venusian</a> · <a href="ymim1" target="_parent">Modeling</a> · <a href="ymihpa.html">Human Practice</a> · <a href="ymijf.html">Extras</a> · <a href="ymit1">Team</a> · <a href="https://2012.igem.org/Main_Page">iGEM</a> </div> </div> <div id="ymi_container_header"> <div id="ymi_header"> <div id="inner_header"> <a href="https://2012.igem.org/Team:NYMU-Taipei"><img src="https://static.igem.org/mediawiki/2012/7/7d/Ymi_header.jpg" border="0"></a> </div> </div> <div class="ymi_spacer_header"></div> </div> <div id="ymi_container"> <div id="ymi_left_column"> <div class="text_area" align="justify"> <div class="title">Protocol</div> <div align="left"> <p><span class="subtitle">J774 Macrophage Cell Culturing</span></p> </div> <div align="left"> <p><img src="images/p1.gif" alt="" width="609" height="442" /></p> <ol> <li>J774 cell line, a murine macrophages cell line established from a tumor that arose in a female BALB/c mouse, were provided by Prof. Yi-Hsuan Lee from National Yang Ming University. </li> <li>View cultures using a microscopy and after the cultures was full of the dish, remove the spent medium. </li> <li>Wash the cell monolayer with PBS and remove it.</li> <li>Add medium(1ml per 25 cm2 of surface area) and scrape cells down.</li> <li>Ensure cells were detached by a microscopy.</li> <li>Centrifuge under 2000 rpm, 5minutes, 25°C.</li> <li>Remove the supernatant and add 5 ml medium to suspend cells.</li> <li>Count the cells by Bright Line Counting Chamber. Subject the cell diversity to 104/ml.</li> <li>Seed the cells for 104/well, 1ml/well in the 24-well plate.</li> <li>Until cells grow full of each well, do the coculturing experiment.<br /> </li></ol></div> </div> </div> <div id="ymi_right_column"> <div class="drawers-wrapper"> <div class="boxcap captop"></div> <ul class="drawers"> <li class="drawer"> <h2 class="drawer-handle">Protocol</h2> <ul> <li><a title="J774 macrophage cell line culturing" href="p1.html">J774 macrophage cell <br /> culturing</a></li> <li><a title="Mesenchymal stem cells culturing" href="p2.html">Mesenchymal stem cells<br /> culturing</a></li> <li><a title="Reprogramming of Somatic Cells into Stem & Separation of iPS cells" href="p3.html">Reprogramming of Somatic<br /> Cells into Stem Cell & <br /> Separation of iPS cells</a></li> <li></li> <li></li> </ul> <h2 class="drawer-handle">Extras</h2> <ul> <li><a title="Judging Form" href="jf.html">Judging Form</a></li> <li><a title="Safety" href="sf.html">Safety</a></li> <li><a title="Collaboration" href="co.html">Collaboration</a></li> <li><a title="Parts" href="parts.html">Parts</a></li> </ul> </li> <li class="drawer last"> <h2 class="drawer-handle">Notebook</h2> <ul> <li><a title="Week 1 ~ Week 4" href="w1.html">Week 1 ~ Week 4</a></li> <li><a title="Week 5 ~ Week 8" href="w2.html">Week 5 ~ Week 8</a></li> <li><a title="Week 9 ~ Week 11" href="w3.html">Week 9 ~ Week 11</a></li> </ul> </li> </ul> <div class="boxcap"></div> </div> </div> <div id="ymi_footer"> </div> </div> <div class="ymi_spacer"></div> </div> </body> </html>