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- <!-- SOP-SC-005-E01-01 人脐带间充质干细胞原代重铺操作记录(A4 竖版) -->
- <div class="sop-record sop-a4 sop-sc-005" 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;">
- <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;">
- <colgroup>
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- <tbody>
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- <td rowspan="2" style="border:1px solid #334155; padding:6px; text-align:center; vertical-align:middle;">
- <img src="/sop-templates/SOP-SC-005-logo.jpeg" alt="logo" width="48" height="48" style="display:block; margin:0 auto;" />
- </td>
- <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;">
- 人脐带间充质干细胞原代重铺操作记录
- </td>
- <td style="border:1px solid #334155; padding:6px 10px; font-size: 12pt; vertical-align:middle;">
- 文件编码: SOP-SC-005-E01
- </td>
- </tr>
- <tr>
- <td style="border:1px solid #334155; padding:6px 10px; font-size: 12pt; vertical-align:middle;">
- 版 本 号: 01
- </td>
- </tr>
- </tbody>
- </table>
- <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;">
- <colgroup>
- <col style="width:8.1%" />
- <col style="width:3.3%" />
- <col style="width:20.3%" />
- <col style="width:1.4%" />
- <col style="width:2.4%" />
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- <col style="width:5.1%" />
- <col style="width:13.7%" />
- </colgroup>
- <tbody><tr class="sop-row-hidden sop-row-hidden">
- <td colspan="6" style="border:1px solid #334155; padding:8px;">工序: 脐带MSC原代重铺操作</td>
- <td colspan="2" style="border:1px solid #334155; padding:8px; white-space:nowrap;">记录编号</td>
- <td colspan="5" class="sop-blank-cell" style="border:1px solid #334155; padding:8px; min-height:28px;"> </td>
- </tr><tr class="sop-row-hidden sop-row-hidden">
- <td colspan="2" style="border:1px solid #334155; padding:8px; white-space:nowrap;">细胞批号</td>
- <td colspan="4" class="sop-blank-cell" style="border:1px solid #334155; padding:8px;"> </td>
- <td colspan="3" style="border:1px solid #334155; padding:8px; white-space:nowrap;">日期</td>
- <td colspan="4" class="sop-blank-cell" style="border:1px solid #334155; padding:8px;"> </td>
- </tr><tr class="sop-row-hidden sop-row-hidden">
- <td rowspan="6" style="border:1px solid #334155; padding:3px 6px; text-align:center; vertical-align:middle; line-height:1.25;">试剂</td>
- <td colspan="2" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">名称</td>
- <td colspan="6" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">生产厂家</td>
- <td colspan="2" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">批号</td>
- <td colspan="2" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">使用数量</td>
- </tr><tr class="sop-row-hidden sop-row-hidden">
- <td colspan="2" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">磷酸盐缓冲液(PBS)</td>
- <td colspan="6" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
- <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
- <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
- </tr><tr class="sop-row-hidden sop-row-hidden">
- <td colspan="2" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">MSC完全培养基</td>
- <td colspan="6" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
- <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
- <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
- </tr><tr class="sop-row-hidden sop-row-hidden">
- <td colspan="2" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">胰酶</td>
- <td colspan="6" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
- <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
- <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
- </tr><tr class="sop-row-hidden sop-row-hidden">
- <td colspan="2" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">生理盐水</td>
- <td colspan="6" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
- <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
- <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
- </tr><tr class="sop-row-hidden sop-row-hidden">
- <td colspan="2" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">消化液</td>
- <td colspan="6" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">------</td>
- <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>
- <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;"> </td>
- </tr><tr class="sop-row-hidden sop-row-hidden">
- <td style="border:1px solid #334155; padding:8px; text-align:center; vertical-align:middle;">仪器<br />设备</td>
- <td colspan="12" style="border:1px solid #334155; padding:8px; vertical-align:top;">
- <div>
- 生物安全柜:
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>XW015</span>
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>XW022</span>
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>其他</span>
- <span class="sop-underline" style="text-align:left; min-width:80px;"> </span>
- </div>
- <div style="margin-top:6px;">
- 离心机:
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>XW016</span>
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>XW026</span>
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>其他</span>
- <span class="sop-underline" style="text-align:left; min-width:80px;"> </span>
- </div>
- </td>
- </tr><tr class="sop-fill-row">
- <td class="sop-fill-label" style="border:1px solid #334155; padding:8px; text-align:center; vertical-align:middle; height:100%;">干细胞原代<br />培养(重铺)</td>
- <td class="sop-fill-cell" colspan="12" style="border:1px solid #334155; padding:10px; vertical-align:top; height:100%;">
- <p style="margin:0 0 6px;">
- 1. 显微镜下观察原代细胞, Day<span class="sop-underline" style="text-align:left; min-width:40px;"> </span>细胞融合率达到90%以上, 进行重铺操作.
- </p>
- <p style="margin:0 0 6px;">
- 2. 在培养瓶表面喷洒酒精后, 用无尘布擦拭放入生物安全柜中, 打开瓶盖, 使用移液管轻轻吸出上清及组织
- <span class="sop-underline" style="text-align:left; min-width:56px;"> </span>mL, 之后
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>是</span>
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>否</span>
- 将上清用于二次贴壁;
- </p>
- <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>
- <p style="margin:0 0 6px;">4. 吸取2.5mL消化液至培养瓶中, 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>
- <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300·g, 离心5分钟, 升9降9;</p>
- <p style="margin:0 0 6px;">
- 6. 倒掉上清, 盖上瓶盖, 轻弹瓶底重悬细胞, 洗管3次, 定容1个50mL离心管, 每管
- <span class="sop-underline" style="text-align:left; min-width:56px;"> </span>mL;
- </p>
- <p style="margin:0 0 6px;">
- 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
- <span class="sop-underline" style="text-align:left; min-width:64px;"> </span>*10<sup>7</sup>个, 接种
- <span class="sop-underline" style="text-align:left; min-width:40px;"> </span>个T175培养瓶, 每瓶120万个细胞;
- </p>
- <p style="margin:0;">8. 充分晃匀后做好重铺P1标记, 放入CO<sub>2</sub>培养箱进行继续培养.</p>
- </td>
- </tr><tr class="sop-row-hidden sop-row-hidden">
- <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>
- <td colspan="6" class="sop-option-cell" style="border:1px solid #334155; padding:8px; text-align:center;">
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>是</span>
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>否</span>
- </td>
- </tr><tr class="sop-row-hidden sop-row-hidden">
- <td rowspan="3" style="border:1px solid #334155; padding:8px; text-align:center; vertical-align:middle;">清场</td>
- <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>
- <td colspan="6" class="sop-option-cell" style="border:1px solid #334155; padding:8px; text-align:center;">
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>是</span>
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>否</span>
- </td>
- </tr><tr class="sop-row-hidden sop-row-hidden">
- <td colspan="6" style="border:1px solid #334155; padding:8px;">废液是否进行收集处理</td>
- <td colspan="6" class="sop-option-cell" style="border:1px solid #334155; padding:8px; text-align:center;">
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>是</span>
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>否</span>
- </td>
- </tr><tr class="sop-row-hidden sop-row-hidden">
- <td colspan="4" style="border:1px solid #334155; padding:8px; white-space:nowrap;">是否清洁, 消毒操作间</td>
- <td colspan="1" class="sop-option-cell" style="border:1px solid #334155; padding:8px; white-space:nowrap; text-align:center;">
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>是</span>
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>否</span>
- </td>
- <td colspan="6" 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>
- <td colspan="1" class="sop-option-cell" style="border:1px solid #334155; padding:8px; white-space:nowrap; text-align:center;">
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>是</span>
- <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false"> </span>否</span>
- </td>
- </tr><tr class="sop-row-hidden sop-row-hidden">
- <td style="border:1px solid #334155; padding:8px; text-align:center;">备注</td>
- <td colspan="12" style="border:1px solid #334155; padding:8px; min-height:48px; height:48px; vertical-align:top;"> </td>
- </tr><tr class="sop-row-hidden sop-row-hidden">
- <td colspan="4" style="border:1px solid #334155; padding:10px;">操作人/日期:</td>
- <td colspan="6" style="border:1px solid #334155; padding:10px;">复核人/日期:</td>
- <td colspan="3" style="border:1px solid #334155; padding:10px;">生产主管/日期:</td>
- </tr></tbody>
- </table>
- </div>
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