SOP-SC-005-E01-01.html 14 KB

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  1. <!-- 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) -->
  2. <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;">
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  12. <img src="/sop-templates/SOP-SC-005-logo.jpeg" alt="logo" width="48" height="48" style="display:block; margin:0 auto;" />
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  15. 人脐带间充质干细胞原代重铺操作记录
  16. </td>
  17. <td style="border:1px solid #334155; padding:6px 10px; font-size: 12pt; vertical-align:middle;">
  18. 文件编码: SOP-SC-005-E01
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  20. </tr>
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  22. <td style="border:1px solid #334155; padding:6px 10px; font-size: 12pt; vertical-align:middle;">
  23. 版 本 号: 01
  24. </td>
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  27. </table>
  28. <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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  44. <tbody><tr class="sop-row-hidden sop-row-hidden">
  45. <td colspan="6" style="border:1px solid #334155; padding:8px;">工序: 脐带MSC原代重铺操作</td>
  46. <td colspan="2" style="border:1px solid #334155; padding:8px; white-space:nowrap;">记录编号</td>
  47. <td colspan="5" class="sop-blank-cell" style="border:1px solid #334155; padding:8px; min-height:28px;">&nbsp;</td>
  48. </tr><tr class="sop-row-hidden sop-row-hidden">
  49. <td colspan="2" style="border:1px solid #334155; padding:8px; white-space:nowrap;">细胞批号</td>
  50. <td colspan="4" class="sop-blank-cell" style="border:1px solid #334155; padding:8px;">&nbsp;</td>
  51. <td colspan="3" style="border:1px solid #334155; padding:8px; white-space:nowrap;">日期</td>
  52. <td colspan="4" class="sop-blank-cell" style="border:1px solid #334155; padding:8px;">&nbsp;</td>
  53. </tr><tr class="sop-row-hidden sop-row-hidden">
  54. <td rowspan="6" style="border:1px solid #334155; padding:3px 6px; text-align:center; vertical-align:middle; line-height:1.25;">试剂</td>
  55. <td colspan="2" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">名称</td>
  56. <td colspan="6" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">生产厂家</td>
  57. <td colspan="2" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">批号</td>
  58. <td colspan="2" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">使用数量</td>
  59. </tr><tr class="sop-row-hidden sop-row-hidden">
  60. <td colspan="2" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">磷酸盐缓冲液(PBS)</td>
  61. <td colspan="6" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">&nbsp;</td>
  62. <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">&nbsp;</td>
  63. <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">&nbsp;</td>
  64. </tr><tr class="sop-row-hidden sop-row-hidden">
  65. <td colspan="2" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">MSC完全培养基</td>
  66. <td colspan="6" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">&nbsp;</td>
  67. <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">&nbsp;</td>
  68. <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">&nbsp;</td>
  69. </tr><tr class="sop-row-hidden sop-row-hidden">
  70. <td colspan="2" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">胰酶</td>
  71. <td colspan="6" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">&nbsp;</td>
  72. <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">&nbsp;</td>
  73. <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">&nbsp;</td>
  74. </tr><tr class="sop-row-hidden sop-row-hidden">
  75. <td colspan="2" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">生理盐水</td>
  76. <td colspan="6" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">&nbsp;</td>
  77. <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">&nbsp;</td>
  78. <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">&nbsp;</td>
  79. </tr><tr class="sop-row-hidden sop-row-hidden">
  80. <td colspan="2" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">消化液</td>
  81. <td colspan="6" style="border:1px solid #334155; padding:3px 6px; text-align:center; line-height:1.25;">------</td>
  82. <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>
  83. <td colspan="2" class="sop-blank-cell" style="border:1px solid #334155; padding:3px 6px; line-height:1.25;">&nbsp;</td>
  84. </tr><tr class="sop-row-hidden sop-row-hidden">
  85. <td style="border:1px solid #334155; padding:8px; text-align:center; vertical-align:middle;">仪器<br />设备</td>
  86. <td colspan="12" style="border:1px solid #334155; padding:8px; vertical-align:top;">
  87. <div>
  88. 生物安全柜:
  89. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>XW015</span>
  90. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>XW022</span>
  91. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>其他</span>
  92. <span class="sop-underline" style="text-align:left; min-width:80px;">&nbsp;</span>
  93. </div>
  94. <div style="margin-top:6px;">
  95. 离心机:
  96. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>XW016</span>
  97. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>XW026</span>
  98. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>其他</span>
  99. <span class="sop-underline" style="text-align:left; min-width:80px;">&nbsp;</span>
  100. </div>
  101. </td>
  102. </tr><tr class="sop-fill-row">
  103. <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>
  104. <td class="sop-fill-cell" colspan="12" style="border:1px solid #334155; padding:10px; vertical-align:top; height:100%;">
  105. <p style="margin:0 0 6px;">
  106. 1. 显微镜下观察原代细胞, Day<span class="sop-underline" style="text-align:left; min-width:40px;">&nbsp;</span>细胞融合率达到90%以上, 进行重铺操作.
  107. </p>
  108. <p style="margin:0 0 6px;">
  109. 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
  110. <span class="sop-underline" style="text-align:left; min-width:56px;">&nbsp;</span>mL, 之后
  111. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>是</span>
  112. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>否</span>
  113. 将上清用于二次贴壁;
  114. </p>
  115. <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>
  116. <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>
  117. <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>
  118. <p style="margin:0 0 6px;">
  119. 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
  120. <span class="sop-underline" style="text-align:left; min-width:56px;">&nbsp;</span>mL;
  121. </p>
  122. <p style="margin:0 0 6px;">
  123. 7. 充分混匀后, 计数<span class="sop-underline" style="text-align:left; min-width:64px;">&nbsp;</span>*10<sup>6</sup>/mL, 共计数量为
  124. <span class="sop-underline" style="text-align:left; min-width:64px;">&nbsp;</span>*10<sup>7</sup>个, 接种
  125. <span class="sop-underline" style="text-align:left; min-width:40px;">&nbsp;</span>个T175培养瓶, 每瓶120万个细胞;
  126. </p>
  127. <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>
  128. </td>
  129. </tr><tr class="sop-row-hidden sop-row-hidden">
  130. <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>
  131. <td colspan="6" class="sop-option-cell" style="border:1px solid #334155; padding:8px; text-align:center;">
  132. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>是</span>
  133. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>否</span>
  134. </td>
  135. </tr><tr class="sop-row-hidden sop-row-hidden">
  136. <td rowspan="3" style="border:1px solid #334155; padding:8px; text-align:center; vertical-align:middle;">清场</td>
  137. <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>
  138. <td colspan="6" class="sop-option-cell" style="border:1px solid #334155; padding:8px; text-align:center;">
  139. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>是</span>
  140. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>否</span>
  141. </td>
  142. </tr><tr class="sop-row-hidden sop-row-hidden">
  143. <td colspan="6" style="border:1px solid #334155; padding:8px;">废液是否进行收集处理</td>
  144. <td colspan="6" class="sop-option-cell" style="border:1px solid #334155; padding:8px; text-align:center;">
  145. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>是</span>
  146. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>否</span>
  147. </td>
  148. </tr><tr class="sop-row-hidden sop-row-hidden">
  149. <td colspan="4" style="border:1px solid #334155; padding:8px; white-space:nowrap;">是否清洁, 消毒操作间</td>
  150. <td colspan="1" class="sop-option-cell" style="border:1px solid #334155; padding:8px; white-space:nowrap; text-align:center;">
  151. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>是</span>
  152. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>否</span>
  153. </td>
  154. <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>
  155. <td colspan="1" class="sop-option-cell" style="border:1px solid #334155; padding:8px; white-space:nowrap; text-align:center;">
  156. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>是</span>
  157. <span class="sop-option"><span class="sop-check" contenteditable="false" data-mce-contenteditable="false">&nbsp;</span>否</span>
  158. </td>
  159. </tr><tr class="sop-row-hidden sop-row-hidden">
  160. <td style="border:1px solid #334155; padding:8px; text-align:center;">备注</td>
  161. <td colspan="12" style="border:1px solid #334155; padding:8px; min-height:48px; height:48px; vertical-align:top;">&nbsp;</td>
  162. </tr><tr class="sop-row-hidden sop-row-hidden">
  163. <td colspan="4" style="border:1px solid #334155; padding:10px;">操作人/日期:</td>
  164. <td colspan="6" style="border:1px solid #334155; padding:10px;">复核人/日期:</td>
  165. <td colspan="3" style="border:1px solid #334155; padding:10px;">生产主管/日期:</td>
  166. </tr></tbody>
  167. </table>
  168. </div>