培育细胞的十个诀窍

<p class="MsoNormal"><span style="background-color: rgb(255, 204, 153);"><span style="font-family: 宋体; font-size: 9pt;">细胞可是许多生物学实验的主角。如果细胞长不好，那么实验的结果也不会好到哪儿去。如何让细胞长得好？</span><span style="font-family: 宋体; font-size: 9pt;">我司</span><span style="font-family: 宋体; font-size: 9pt;">在此介绍了让细胞培育的十个诀窍。</span></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="color: rgb(255, 0, 0);"><strong><span style="font-family: 宋体; font-size: 9pt;">1.</span></strong><span style="color: rgb(0, 0, 0);"><strong><span style="font-family: 宋体; font-size: 9pt;"> </span></strong><span style="font-family: 宋体; font-size: 9pt;"><font face="宋体">确保所有实验室材料都无菌</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">交叉污染是细胞培养的大敌。即使是轻微的污染，也可能毁了几个星期的成果。因培养箱内温暖潮湿，真菌极易生长，因此必须注意定期清洁。此外，在将培养瓶、移液管及其他的相关物品放入超净台之前，应用酒精擦拭干净，以避免污染。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="color: rgb(255, 0, 0);"><strong><span style="font-family: 宋体; font-size: 9pt;">2. </span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋体; font-size: 9pt;"><font face="宋体">小心处理您的培养物</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋体">细胞培养的脆弱性怎么强调也不为过。剧烈摇晃，或连续的温度波动可能会对生长产生不利的影响。确保培养箱是水平的，温度均一，且远离电动仪器。此外，尽量避免一次处理多个细胞系，因为它可能会影响细胞的基因型和表型。您还应定期完成</font>STR<font face="宋体">图谱分析，以确认细胞系的身份。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="color: rgb(255, 0, 0);"><strong><span style="font-family: 宋体; font-size: 9pt;">3.</span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋体; font-size: 9pt;"> <font face="宋体">在使用前正确解冻细胞</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋体">尽管解冻看起来是个简单的步骤，但必须操作得当，以免伤害细胞。长时间暴露在高温条件下会使细胞无法铺板。因此，冻存管应当在</font>37<font face="宋体">&deg;</font><font face="Calibri">C</font><font face="宋体">水浴中放置</font><font face="Calibri">2</font><font face="宋体">分钟，然后用条件培养基稀释，以避免造成直接伤害。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="color: rgb(255, 0, 0);"><strong><span style="font-family: 宋体; font-size: 9pt;">4. </span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋体; font-size: 9pt;"><font face="宋体">使用对数生长期的细胞</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋体">细胞培养过程共有</font>3<font face="宋体">个阶段：停滞期、对数期和平台期，分别代表了低细胞生长、高细胞生长和无细胞生长。有活力的细胞是健康的，快速分裂的，在对数期以</font><font face="Calibri">70-80%</font><font face="宋体">的汇合度存在。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="color: rgb(255, 0, 0);"><strong><span style="font-family: 宋体; font-size: 9pt;">5. </span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋体; font-size: 9pt;"><font face="宋体">在传代之前不要让细胞完全汇合</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><font face="宋体">汇合度是指贴壁细胞占据培养瓶表面积的比例。完全汇合意味着</font>100%<font face="宋体">的表面都被贴壁细胞覆盖。一定要避免这种状态，因为它意味着细胞不能继续生长。不过，当务之急是使用活跃生长的细胞。</font></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="color: rgb(255, 0, 0);"><strong><span style="font-family: 宋体; font-size: 9pt;">6.</span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋体; font-size: 9pt;"> <font face="宋体">选择培养基开发策略</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">在细胞培养过程中，培养基是控制产品质量、产量和成本的重要因素。必须针对每种培养物来定制培养基，以优化实验结果。在决定以哪种方式来开发您的培养基时，您有几个选择。您可以购买现成的，自己开发，也可以与另一家公司合作开发特定的培养基。在这个过程中，您需要考虑时间和成本等因素。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="color: rgb(255, 0, 0);"><strong><span style="font-family: 宋体; font-size: 9pt;">7. </span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋体; font-size: 9pt;"><font face="宋体">配制干粉培养基时评估水的质量</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">液体培养基往往会带来比干粉培养基更高质量的结果。这可能与水的质量有关。由于细菌在水中迅速生长，故需要监控内毒素及其他污染物。商业公司有资源来监控和控制细菌生长，比如过滤器。因此使用商业化生产的液体培养基会更加可靠一点。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="color: rgb(255, 0, 0);"><strong><span style="font-family: 宋体; font-size: 9pt;">8. </span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋体; font-size: 9pt;"><font face="宋体">利用细胞的代谢特性来优化培养基</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">分析使用过的培养基，有助于鉴定必要成分的代谢速率，包括氨基酸和维生素。从细胞生长阶段和蛋白生产阶段收集到的动态代谢图谱可用来平衡基础培养基和添加培养基，指导培养基的重点，有助于蛋白质的生产。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="color: rgb(255, 0, 0);"><strong><span style="font-family: 宋体; font-size: 9pt;">9.</span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋体; font-size: 9pt;"> <font face="宋体">避免细胞的过度消化</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">胰蛋白酶通过消化负责让细胞与容器结合的蛋白质，而实现贴壁细胞的传代。如果细胞暴露在胰酶中时间太长，则胰酶将开始切割细胞表面蛋白，这会影响细胞行驶正常功能的能力。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="color: rgb(255, 0, 0);"><strong><span style="font-family: 宋体; font-size: 9pt;">10. </span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋体; font-size: 9pt;"><font face="宋体">培养基中不要一直使用抗生素</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p> <p class="MsoNormal"><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;">随着细胞培养物更加频繁地暴露在抗生素中，对抗生素有着天然耐药性的菌株将开始出现。这种现象可掩盖潜在的污染，这对实验是非常有害的。</span><span style="mso-spacerun:'yes';font-family:宋体;mso-ascii-font-family:Calibri;mso-hansi-font-family:Calibri;mso-bidi-font-family:'Times New Roman';font-size:9.0000pt;mso-font-kerning:1.0000pt;"><o:p></o:p></span></p>