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常用细胞房操作
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==更换培养基== #更换频率1次/d。 #弃去旧培养基: ##贴壁细胞(如癌细胞):移液枪直接吸取培养基并弃去; ##悬浮细胞:收集原培养基及cell置于EP管中,离心,弃上清: ##半贴壁细胞:采用半换液,倾斜孔板或培养皿,移液枪吸取上层培养基弃去。 #无需PBS清洗,直接加入新鲜培养基。培养基中的酚红用于判断培养基有无过期。培养基总量:10cm 培养皿6-10m。 <table class="wikitable" style="margin: auto; border-collapse: collapse; border: 1px solid #a2a9b1;"> <caption>表1:贴壁细胞或悬浮细胞接种数量、培养基体积</caption> <tr> <th>培养器皿</th> <th>每孔表面积<br>(cm²)</th> <th>每孔培养基的<br>体积(ml)</th> <th>贴壁细胞转染前一天<br>接种密度</th> <th>悬浮细胞转染当<br>天接种密度</th> </tr> <tr> <td>96孔板</td> <td>0.3</td> <td>0.1</td> <td>5000±2500</td> <td>2-5×10<sup>5</sup></td> </tr> <tr> <td>24孔板</td> <td>2</td> <td>0.5</td> <td>25000±10000</td> <td>1-2.5×10<sup>5</sup></td> </tr> <tr> <td>12孔板</td> <td>4</td> <td>1</td> <td>50000±20000</td> <td>2-5×10<sup>5</sup></td> </tr> <tr> <td>6孔板</td> <td>10</td> <td>2</td> <td>150000±50000</td> <td>0.4-1×10<sup>6</sup></td> </tr> <tr> <td>60mm培养皿</td> <td>20</td> <td>4</td> <td>400000±100000</td> <td>1-2.5×10<sup>6</sup></td> </tr> <tr> <td>100mm培养皿</td> <td>60</td> <td>10</td> <td>1×10<sup>6</sup>±250000</td> <td>2-5×10<sup>6</sup></td> </tr> <tr style="border-top: 1px solid #a2a9b1;"> <td colspan="2"><i>*:乙醇待水溶液体系完全溶解后再加入</i></td> <td colspan="2"><i>*:贴壁细胞培养密度取决于培养器皿的表面积而悬浮细胞则由培养基的体积决定。 </tr> </table>
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