基因改造的人类介质细胞带有微载体的可扩展制造在旋转瓶中
Pedro Silva Couto1, Dale J Stibbs1, Marco C Rotondi1
1Department of Biochemical Engineering, Advanced Centre for Biochemical Engineering, University College London, Gower Street, London, WC1E 6BT, UK.
Applied microbiology and biotechnology
|July 20, 2023
概括
用微载体生物反应器系统实现了基因改造的人类介质细胞 (hMSCs) 的可扩展制造. 这一过程成功地扩大了基因工程hMSCs,而不会影响其临床应用的基本特征.
科学领域:
- 细胞生物学 细胞生物学
- 生物技术是生物技术.
- 再生医学是一种再生医学.
背景情况:
- 人类介质干细胞 (hMSCs) 具有免疫调节特性和分化潜力,导致了1000多项临床试验.
- 基因改造的hMSC产品正在进入临床试验,需要可扩展的制造解决方案.
研究的目的:
- 使用微载体生物反应器系统开发和演示基因改造的hMSC可扩展的制造工艺.
- 评估lentiviral转导和扩张对hMSC免疫表型和细胞度的影响.
主要方法:
- 带组织中介细胞 (UCT-hMSCs) 被转化为编码GFP或VEGF的lentiviral载体.
- 细胞在旋转瓶中的Spherecol微载体上扩展了七天.
- 分析了细胞度,葡萄糖/乳酸盐水平以及免疫表型标记 (CD73,CD90,CD105,CD11b,CD19,CD34,CD45,HLA-DR) 的情况.
主要成果:
- 对于非转和转的hMSCs,都实现了可比的细胞度 (≈1.8 × 10^5细胞/毫升).
- 观察到葡萄糖消耗和乳酸积累,这是细胞扩张的典型特征.
- 免疫类型分析证实,基因修饰和扩展维持了hMSC表面受体表达率高于90%,负标记低于5%,符合ISCT标准.
结论:
- 一个可扩展的微载体生物反应器系统可以有效扩展基因改造的hMSCs.
- 制造过程保留了hMSCs的关键免疫类型特征.
- 这种方法解决了基因改造的hMSC疗法的临床转化和商业化方面的重大瓶.
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