不同条件对肿瘤球制备单细胞效果的评价

doi:10.3969/j.issn.1000-1158.2025.02.19

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Abstract The choice of single-cell isolation and preparation techniques can significantly impact cell status and physiological functions， ultimately leading to biases in related studies. This study employs a tumor spheroid model with strong cultivation consistency and utilizes four separation methods， including water bath， metal bath， mechanical， and optimized water bath methods， along with three types of dissociation enzymes—collagenase， trypsin， and a human tumor gentle enzyme kit—to isolate multiple single-cell sample groups. Key indicators such as single-cell quantity， viability， metabolic products， and drug pathways were investigated. The results indicate that the optimized water bath method yields the highest single-cell viability at （90.27±8.56）%， while trypsin demonstrates the best performance among the enzymes， with a single-cell viability of （93.20±3.84）% and the highest cell yield. Collagenase-prepared single cells exhibit the strongest clonal proliferation ability and express fewer metabolic changes compared to the tumor spheroid， closely resembling the true state of the tumor spheroid. Trypsin-prepared single cells show downregulation of metabolites involved in methionine metabolism， reducing the sensitivity of cancer cells to drugs and affecting the assessment of drug efficacy.

Key words： biometrology tumorsphere single-cell preparation mass spectrometry metabolom

Received: 20 March 2024 Published: 04 March 2025

PACS:

TB99

Fund:The basic scientific research expenses of the Chinese Academy of Metrology

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	Articles by authors
	YANG Lulu
	ZHANG Yongzhuo
	GONG Xiaoyun
	XUE Zhichao
	SHENTU Xuping

Cite this article:

YANG Lulu,ZHANG Yongzhuo,GONG Xiaoyun, et al. Evaluation of the Effect of Different Conditions on Single Cell Preparation of Tumorsphere[J]. Acta Metrologica Sinica, 2025, 46(2): 298-308.

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http://jlxb.china-csm.org:81/Jwk_jlxb/EN/10.3969/j.issn.1000-1158.2025.02.19 OR http://jlxb.china-csm.org:81/Jwk_jlxb/EN/Y2025/V46/I2/298

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