| 杨栋1，2 牛红军3 陆刚2 史嘉林2 孙浩明2 李晖1△.悬浮培养CHO 细胞生产重组人促红细胞生成素条件的优化*[J].,2012,12(26):5053-5056 |
| 悬浮培养CHO 细胞生产重组人促红细胞生成素条件的优化* |
| An Optimized Method for Suspension Culture of CHO Cells to ProduceRecombinant Human Erythropoietin (EPO)* |
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| DOI： |
| 中文关键词: 重组人促红细胞生成素 悬浮培养 贴壁培养 |
| 英文关键词: Recombinant human erythropoietin (EPO) Suspension culture Adherent culture |
| 基金项目:国家自然科学基金面上项目（30771863）；国家自然科学基金面上项目（81172616）；
哈尔滨科技厅优秀学科带头人基金项目（RC2008XK004039） |
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| 中文摘要: |
| 目的：通过对贴壁培养CHO 细胞筛选驯化，得到高表达的细胞后进行悬浮培养生产重组人促红细胞生成素（rHuEPO）。方
法：利用96孔板和24孔板对CHO细胞进行筛选，得到高表达细胞株后进行驯化，使其适合悬浮培养，经过摇瓶扩增后接种到生
物反应器中无血清培养，每天监测葡萄糖含量，测rHuEPO表达量。结果：悬浮培养CHO 细胞生产rHuEPO，生产周期短，表达量
比贴壁培养高出很多，操作方便，减少污染，易于放大，并建立了适合悬浮培养的CHO 细胞株，为工业化悬浮培养CHO 细胞生产
rHuEPO提供了技术基础。结论：经过工艺优化后利用无血清悬浮培养生产促红细胞生成素平均表达量较贴壁培养高，生产周期
短，有利于降低生产成本。 |
| 英文摘要: |
| Objective: To screen and domesticate the adherent cultured CHO cells to obtain high expression of cell suspension
culture for production of recombinant human erythropoietin erythropoietin (rHuEPO). Methods: Using 96-well and 24-well plates culture
method to screen and domesticate the highly expressing CHO cell strain. Acclimate the high expression cell strain and make it suitable
for suspension culture. It's inoculated into the bioreactor in serum-free culture after amplified by the shake flask, and monitoring of
glucose content, measuring rHuEPO expression of daily. Results: The suspension culture of CHO cell production of rHuEPO has short
production period, higher expression than adherent culture. On the other hand, it is easy to operate and scale-up, but not easy to pollute.
Furthermore, we established of the CHO cell strain for suspension culture,which provided a technical basis for industrial production of
CHO cells the rHuEPO. Conclusion: After process optimization, the use of serum-free suspension culture production of erythropoietin
average expression has high, short production period, low cost of production.than adherent culture. |
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