183 lines
15 KiB
HTML
183 lines
15 KiB
HTML
<!-- SOP-SC-007-E01-01 人脐带间充质干细胞传代培养操作记录(A4 竖版) -->
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<div class="sop-record sop-a4 sop-sc-007" style="font-family: 'PingFang SC','Microsoft YaHei',sans-serif; font-size: 10.5pt; color: #1e293b; line-height: 1.55; box-sizing: border-box; display: flex; flex-direction: column; height: 297mm; min-height: 297mm; padding: 16mm 15mm 20mm;">
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<table class="sop-header-table sop-row-hidden" style="width:100%; border-collapse:collapse; table-layout:fixed; border:1px solid #334155; margin-bottom:8px; flex-shrink:0;">
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<colgroup>
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<col style="width:15%" />
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<col style="width:50%" />
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<col style="width:35%" />
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</colgroup>
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<tbody>
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<tr>
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<td rowspan="2" style="border:1px solid #334155; padding:6px; text-align:center; vertical-align:middle;">
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<img src="/sop-templates/SOP-SC-007-logo.jpeg" alt="logo" width="48" height="48" style="display:block; margin:0 auto;" />
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</td>
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<td rowspan="2" style="border:1px solid #334155; padding:8px; text-align:center; vertical-align:middle; font-family: SimSun, 'Songti SC', serif; font-size: 11pt; font-weight:700;">
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人脐带间充质干细胞传代培养操作记录
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</td>
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<td style="border:1px solid #334155; padding:6px 10px; font-size: 12pt; vertical-align:middle;">
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文件编码: SOP-SC-007-E01
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</td>
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</tr>
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<tr>
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<td style="border:1px solid #334155; padding:6px 10px; font-size: 12pt; vertical-align:middle;">
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版 本 号: 01
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</td>
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</tr>
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</tbody>
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</table>
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<table class="sop-body-table" style="width:100%; border-collapse:collapse; table-layout:fixed; border:1px solid #334155; flex:1 1 auto; height:100%; min-height:0;">
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<colgroup>
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<col style="width:8.1%" />
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<col style="width:3.3%" />
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<col style="width:20.2%" />
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<col style="width:1.4%" />
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<col style="width:2.4%" />
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<col style="width:14.7%" />
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<col style="width:5.8%" />
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<col style="width:5.6%" />
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<col style="width:2.5%" />
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<col style="width:17.1%" />
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<col style="width:5.2%" />
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<col style="width:13.7%" />
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</colgroup>
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<tbody><tr class="sop-row-hidden sop-row-hidden">
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<td colspan="6" style="border:1px solid #334155; padding:8px;">工序: 脐带MSC传代操作</td>
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<td colspan="2" style="border:1px solid #334155; padding:8px; white-space:nowrap;">记录编号</td>
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<td colspan="4" class="sop-blank-cell" style="border:1px solid #334155; padding:8px; min-height:28px;"> </td>
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</tr><tr class="sop-row-hidden sop-row-hidden">
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<td colspan="2" style="border:1px solid #334155; padding:8px; white-space:nowrap;">细胞批号</td>
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<td colspan="4" class="sop-blank-cell" style="border:1px solid #334155; padding:8px;"> </td>
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<td colspan="2" style="border:1px solid #334155; padding:8px; white-space:nowrap;">日期</td>
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<td colspan="4" class="sop-blank-cell" style="border:1px solid #334155; padding:8px;"> </td>
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</tr><tr class="sop-row-hidden sop-row-hidden">
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<td rowspan="6" style="border:1px solid #334155; padding:3px 6px; text-align:center; vertical-align:middle; line-height:1.25;">试剂</td>
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<td colspan="2" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">名称</td>
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<td colspan="5" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">生产厂家</td>
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<td colspan="2" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">批号</td>
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<td colspan="2" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">使用数量</td>
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</tr><tr class="sop-row-hidden sop-row-hidden">
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<td colspan="2" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">磷酸盐缓冲液(PBS)</td>
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<td colspan="5" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
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<td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
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<td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
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</tr><tr class="sop-row-hidden sop-row-hidden">
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<td colspan="2" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">MSC完全培养基</td>
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<td colspan="5" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
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<td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
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<td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
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</tr><tr class="sop-row-hidden sop-row-hidden">
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<td colspan="2" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">生理盐水</td>
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<td colspan="5" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
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<td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
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<td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
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</tr><tr class="sop-row-hidden sop-row-hidden">
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<td colspan="2" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">胰酶</td>
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<td colspan="5" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
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<td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
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<td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
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</tr><tr class="sop-row-hidden sop-row-hidden">
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<td colspan="2" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">消化液</td>
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<td colspan="5" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">------</td>
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<td colspan="2" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">制定配制记录</td>
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<td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
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</tr><tr class="sop-row-hidden sop-row-hidden">
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<td style="border:1px solid #334155; padding:8px; text-align:center; vertical-align:middle;">仪器<br />设备</td>
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<td colspan="11" style="border:1px solid #334155; padding:8px; vertical-align:top;">
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<div>
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生物安全柜:
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>XW015</span>
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>XW022</span>
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>其他</span>
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<span class="sop-underline" style="text-align:left; min-width:80px;"> </span>
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</div>
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<div style="margin-top:6px;">
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离心机:
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>XW016</span>
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>XW026</span>
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>其他</span>
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<span class="sop-underline" style="text-align:left; min-width:80px;"> </span>
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</div>
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</td>
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</tr><tr class="sop-fill-row">
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<td class="sop-fill-label" style="border:1px solid #334155; padding:8px; text-align:center; vertical-align:middle; height:100%;">人脐带间充质<br />干细胞<br />传代操作</td>
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<td class="sop-fill-cell" colspan="11" style="border:1px solid #334155; padding:10px; vertical-align:top; height:100%;">
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<p style="margin:0 0 6px;">
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1. 显微镜下细胞综合融合率达到90%以上, 进行传代操作, 此次共处理
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<span class="sop-underline" style="text-align:left; min-width:40px;"> </span>瓶细胞;
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</p>
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<p style="margin:0 0 6px;">
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2. 使用移液管轻轻吸出上清共
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<span class="sop-underline" style="text-align:left; min-width:56px;"> </span>mL于离心管中作为终止消化液; 其余上清共
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<span class="sop-underline" style="text-align:left; min-width:56px;"> </span>mL, 用作因子回收, 做好标记并放入-80℃冰箱;
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</p>
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<p style="margin:0 0 6px;">3. 吸取10mL生理盐水至培养瓶中, 润洗细胞生长面, 再吸弃上清, 重复一次;</p>
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<p style="margin:0 0 6px;">4. 每瓶加入2.5mL消化液, 消化液铺满整瓶后, 在CO<sub>2</sub>培养箱中消化1min, 待在显微镜下观察细胞消化彻底后, 每瓶加入终止消化液10mL终止消化;</p>
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<p style="margin:0 0 6px;">5. 吸出液体收集于离心瓶中, 每瓶加入10mL生理盐水洗涤细胞瓶, 冲洗2次并将所有液体收集至同一离心瓶中, 用离心机400·g, 离心5分钟, 升9降9;</p>
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<p style="margin:0 0 6px;">
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6. 共收获
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>50mL</span>
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>250mL</span>
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离心瓶细胞<span class="sop-underline" style="text-align:left; min-width:40px;"> </span>瓶, 倒掉上清, 盖上瓶盖, 轻弹瓶底重悬细胞, 合管细胞, 并重悬, 洗管
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<span class="sop-underline" style="text-align:left; min-width:40px;"> </span>次, 预估数量后定容
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<span class="sop-underline" style="text-align:left; min-width:40px;"> </span>个50mL离心管, 每管
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<span class="sop-underline" style="text-align:left; min-width:56px;"> </span>mL;
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</p>
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<p style="margin:0 0 6px;">
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7. 充分混匀后, 计数<span class="sop-underline" style="text-align:left; min-width:64px;"> </span>*10<sup>6</sup>/mL, 共计数量为
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<span class="sop-underline" style="text-align:left; min-width:64px;"> </span>*10<sup>7</sup>个, 分出接种传代细胞
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<span class="sop-underline" style="text-align:left; min-width:56px;"> </span>mL, 冻存细胞
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<span class="sop-underline" style="text-align:left; min-width:56px;"> </span>mL; 接种
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<span class="sop-underline" style="text-align:left; min-width:40px;"> </span>个T175培养瓶, 每瓶140万个细胞, 冻存细胞
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<span class="sop-underline" style="text-align:left; min-width:56px;"> </span>*10<sup>7</sup>个,
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<span class="sop-underline" style="text-align:left; min-width:40px;"> </span>管;
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</p>
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<p style="margin:0 0 6px;">8. 配平后, 用离心机400·g, 离心5分钟, 升9降9;</p>
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<p style="margin:0;">
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9. 接种的细胞弃掉上清后, 用MSC完全培养基重悬定容至
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<span class="sop-underline" style="text-align:left; min-width:56px;"> </span>mL, 每个培养瓶接种
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<span class="sop-underline" style="text-align:left; min-width:56px;"> </span>mL, 加入培养基
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<span class="sop-underline" style="text-align:left; min-width:56px;"> </span>mL.
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</p>
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</td>
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</tr><tr class="sop-row-hidden sop-row-hidden">
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<td colspan="7" style="border:1px solid #334155; padding:8px;">每个培养瓶上皆清楚标示细胞批号</td>
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<td colspan="5" class="sop-option-cell" style="border:1px solid #334155; padding:8px; text-align:center;">
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>是</span>
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>否</span>
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</td>
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</tr><tr class="sop-row-hidden sop-row-hidden">
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<td rowspan="3" style="border:1px solid #334155; padding:8px; text-align:center; vertical-align:middle;">清场</td>
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<td colspan="6" style="border:1px solid #334155; padding:8px;">未使用完试剂是否放入指定位置</td>
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<td colspan="5" class="sop-option-cell" style="border:1px solid #334155; padding:8px; text-align:center;">
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>是</span>
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>否</span>
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</td>
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</tr><tr class="sop-row-hidden sop-row-hidden">
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<td colspan="6" style="border:1px solid #334155; padding:8px;">废液是否进行收集处理</td>
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<td colspan="5" class="sop-option-cell" style="border:1px solid #334155; padding:8px; text-align:center;">
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>是</span>
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>否</span>
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</td>
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</tr><tr class="sop-row-hidden sop-row-hidden">
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<td colspan="4" style="border:1px solid #334155; padding:8px; white-space:nowrap;">是否清洁, 消毒操作间</td>
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<td colspan="1" class="sop-option-cell" style="border:1px solid #334155; padding:8px; white-space:nowrap; text-align:center;">
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>是</span>
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>否</span>
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</td>
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<td colspan="5" style="border:1px solid #334155; padding:8px; white-space:nowrap;">是否清洁, 消毒生物安全柜与移液枪</td>
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<td colspan="1" class="sop-option-cell" style="border:1px solid #334155; padding:8px; white-space:nowrap; text-align:center;">
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>是</span>
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<span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>否</span>
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</td>
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</tr><tr class="sop-row-hidden sop-row-hidden">
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<td style="border:1px solid #334155; padding:8px; text-align:center;">备注</td>
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<td colspan="11" style="border:1px solid #334155; padding:8px; min-height:48px; height:48px; vertical-align:top;"> </td>
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</tr><tr class="sop-row-hidden sop-row-hidden">
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<td colspan="4" style="border:1px solid #334155; padding:10px;">操作人/日期:</td>
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<td colspan="5" style="border:1px solid #334155; padding:10px;">复核人/日期:</td>
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<td colspan="3" style="border:1px solid #334155; padding:10px;">生产主管/日期:</td>
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</tr></tbody>
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</table>
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</div>
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