培育細胞的十個訣竅

<p class="MsoNormal"><span style="background-color: rgb(255, 204, 153);"><span style="font-family: 宋體; font-size: 9pt;">細胞可是許多生物學實驗的主角。如果細胞長不好，那么實驗的結果也不會好到哪兒去。如何讓細胞長得好？</span><span style="font-family: 宋體; font-size: 9pt;">我司</span><span style="font-family: 宋體; font-size: 9pt;">在此介紹了讓細胞培育的十個訣竅。</span></span><span style="mso-spacerun:'yes';font-family:宋體;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: 宋體; font-size: 9pt;">1.</span></strong><span style="color: rgb(0, 0, 0);"><strong><span style="font-family: 宋體; font-size: 9pt;"> </span></strong><span style="font-family: 宋體; font-size: 9pt;"><font face="宋體">確保所有實驗室材料都無菌</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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;">交叉污染是細胞培養的大敵。即使是輕微的污染，也可能毀了幾個星期的成果。因培養箱內溫暖潮濕，真菌極易生長，因此必須注意定期清潔。此外，在將培養瓶、移液管及其他的相關物品放入超凈臺之前，應用酒精擦拭干凈，以避免污染。</span><span style="mso-spacerun:'yes';font-family:宋體;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: 宋體; font-size: 9pt;">2. </span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋體; font-size: 9pt;"><font face="宋體">小心處理您的培養物</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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="宋體">細胞培養的脆弱性怎么強調也不為過。劇烈搖晃，或連續的溫度波動可能會對生長產生不利的影響。確保培養箱是水平的，溫度均一，且遠離電動儀器。此外，盡量避免一次處理多個細胞系，因為它可能會影響細胞的基因型和表型。您還應定期完成</font>STR<font face="宋體">圖譜分析，以確認細胞系的身份。</font></span><span style="mso-spacerun:'yes';font-family:宋體;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: 宋體; font-size: 9pt;">3.</span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋體; font-size: 9pt;"> <font face="宋體">在使用前正確解凍細胞</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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="宋體">盡管解凍看起來是個簡單的步驟，但必須操作得當，以免傷害細胞。長時間暴露在高溫條件下會使細胞無法鋪板。因此，凍存管應當在</font>37<font face="宋體">&deg;</font><font face="Calibri">C</font><font face="宋體">水浴中放置</font><font face="Calibri">2</font><font face="宋體">分鐘，然后用條件培養基稀釋，以避免造成直接傷害。</font></span><span style="mso-spacerun:'yes';font-family:宋體;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: 宋體; font-size: 9pt;">4. </span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋體; font-size: 9pt;"><font face="宋體">使用對數生長期的細胞</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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="宋體">細胞培養過程共有</font>3<font face="宋體">個階段：停滯期、對數期和平臺期，分別代表了低細胞生長、高細胞生長和無細胞生長。有活力的細胞是健康的，快速分裂的，在對數期以</font><font face="Calibri">70-80%</font><font face="宋體">的匯合度存在。</font></span><span style="mso-spacerun:'yes';font-family:宋體;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: 宋體; font-size: 9pt;">5. </span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋體; font-size: 9pt;"><font face="宋體">在傳代之前不要讓細胞完全匯合</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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="宋體">匯合度是指貼壁細胞占據培養瓶表面積的比例。完全匯合意味著</font>100%<font face="宋體">的表面都被貼壁細胞覆蓋。一定要避免這種狀態，因為它意味著細胞不能繼續生長。不過，當務之急是使用活躍生長的細胞。</font></span><span style="mso-spacerun:'yes';font-family:宋體;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: 宋體; font-size: 9pt;">6.</span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋體; font-size: 9pt;"> <font face="宋體">選擇培養基開發策略</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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;">在細胞培養過程中，培養基是控制產品質量、產量和成本的重要因素。必須針對每種培養物來定制培養基，以優化實驗結果。在決定以哪種方式來開發您的培養基時，您有幾個選擇。您可以購買現成的，自己開發，也可以與另一家公司合作開發特定的培養基。在這個過程中，您需要考慮時間和成本等因素。</span><span style="mso-spacerun:'yes';font-family:宋體;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: 宋體; font-size: 9pt;">7. </span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋體; font-size: 9pt;"><font face="宋體">配制干粉培養基時評估水的質量</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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;">液體培養基往往會帶來比干粉培養基更高質量的結果。這可能與水的質量有關。由于細菌在水中迅速生長，故需要監控內毒素及其他污染物。商業公司有資源來監控和控制細菌生長，比如過濾器。因此使用商業化生產的液體培養基會更加可靠一點。</span><span style="mso-spacerun:'yes';font-family:宋體;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: 宋體; font-size: 9pt;">8. </span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋體; font-size: 9pt;"><font face="宋體">利用細胞的代謝特性來優化培養基</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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;">分析使用過的培養基，有助于鑒定必要成分的代謝速率，包括氨基酸和維生素。從細胞生長階段和蛋白生產階段收集到的動態代謝圖譜可用來平衡基礎培養基和添加培養基，指導培養基的重點，有助于蛋白質的生產。</span><span style="mso-spacerun:'yes';font-family:宋體;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: 宋體; font-size: 9pt;">9.</span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋體; font-size: 9pt;"> <font face="宋體">避免細胞的過度消化</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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;">胰蛋白酶通過消化負責讓細胞與容器結合的蛋白質，而實現貼壁細胞的傳代。如果細胞暴露在胰酶中時間太長，則胰酶將開始切割細胞表面蛋白，這會影響細胞行駛正常功能的能力。</span><span style="mso-spacerun:'yes';font-family:宋體;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: 宋體; font-size: 9pt;">10. </span></strong><span style="color: rgb(0, 0, 0);"><span style="font-family: 宋體; font-size: 9pt;"><font face="宋體">培養基中不要一直使用抗生素</font></span></span></span><span style="mso-spacerun:'yes';font-family:宋體;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:宋體;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;">隨著細胞培養物更加頻繁地暴露在抗生素中，對抗生素有著天然耐藥性的菌株將開始出現。這種現象可掩蓋潛在的污染，這對實驗是非常有害的。</span><span style="mso-spacerun:'yes';font-family:宋體;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>