单细胞RNA测序揭示了基因表达异质性在不稳定,克隆衍生的CHO细胞系中基于人口的变化
Theodore W Peters1, Jake T Leinas1, Sofie O'Brien1
1Molecular Cellular Technologies, Biotherapeutic Pharm. Sci., Pfizer Inc., Bothell, Washington, USA.
Biotechnology progress
|March 9, 2026
概括
为蛋白质生产而设计的中国仓鼠卵巢细胞系往往会随着时间的推移而失去表达. 这项研究表明,基因表达的细胞异质性增加有助于细胞系制造中的这种不稳定性.
科学领域:
- 生物技术是生物技术.
- 细胞生物学 细胞生物学
- 基因组学就是基因组学.
背景情况:
- 中国仓鼠卵巢 (CHO) 细胞对于产生外来蛋白质至关重要,但与长期表达稳定性作斗争.
- 细胞系不稳定性,以生产力损失为特征,需要选一致的表达.
- 克隆群体内的表型异质性是一种可疑的,但尚未研究的不稳定性原因.
研究的目的:
- 研究细胞异质性在单克隆抗体生产过程中CHO细胞系的不稳定性中的作用.
- 为了比较稳定和不稳定的CHO细胞系在其制造寿命期间的基因表达特征和细胞异质性.
主要方法:
- 单细胞RNA测序在稳定和不稳定的CHO细胞系上进行,这些细胞系产生相同的单克隆抗体.
- 在制造过程中,在早期和晚期进行了分析.
- 使用分子方法来确认基因表达变化和通路激活.
主要成果:
- 与稳定的细胞系相比,不稳定的细胞系表现出明显更高的表达异质性和表达特征集群的动态转移.
- 不稳定的细胞系中差异表达的基因表明随着时间的推移,代谢途径活性发生了变化.
- 分子分析证实了基因表达的变化和与不稳定细胞中受挑战的蛋白质平衡相关的途径的丰富.
结论:
- 克隆性CHO细胞群体内的异质性增加是导致细胞系不稳定的关键因素.
- 了解和管理细胞异质性可以为改善生物制造中的长期表达稳定性提供生物学背景.
- 这项研究为进一步研究工程细胞系中缓解不稳定性的研究提供了基础.
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