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On-line Access: 2023-10-18

Received: 2022-11-27

Revision Accepted: 2023-03-28

Crosschecked: 2023-10-19

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Clicked: 736

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Ling YU

https://orcid.org/0000-0002-6726-281X

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Journal of Zhejiang University SCIENCE A 2023 Vol.24 No.10 P.875-885

http://doi.org/10.1631/jzus.A22D0235


Using pipette tips to readily generate spheroids comprising single or multiple cell types


Author(s):  Rong PAN, Xiaoyan YANG, Shiming WU, Yuanyuan XIE, Feng CHEN, Ke NING, Wei SUN, Ling YU

Affiliation(s):  Key Laboratory of Luminescence Analysis and Molecular Sensing, Ministry of Education, Institute for Clean Energy and Advanced Materials, School of Materials and Energy, Southwest University, Chongqing 400715, China; more

Corresponding email(s):   lingyu12@swu.edu.cn

Key Words:  Pipette tip, 3D cell culture, Tumor spheroids, Co-culture, In-situ observation


Rong PAN, Xiaoyan YANG, Shiming WU, Yuanyuan XIE, Feng CHEN, Ke NING, Wei SUN, Ling YU. Using pipette tips to readily generate spheroids comprising single or multiple cell types[J]. Journal of Zhejiang University Science A, 2023, 24(10): 875-885.

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pages="875-885",
year="2023",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.A22D0235"
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Abstract: 
Three-dimensional (3D) cell culture methods have been validated that can replicate the tumor environment in vivo to a large extent, providing an effective tool for studying tumors. In this study, we demonstrated the use of standard laboratory pipette tips as micro vessels for generating 3D cell spheroids. No microfabrication or wet-chemistry surface modifications were involved in the procedure. Spheroids consisting of single or multiple cell types were generated within 24 h just by pipetting and incubating a cell suspension in pipette tips. Scanning electron microscope and optical microscope proved that the cells grew together tightly, and suggested that while gravity force might have initiated the sedimentation of cells at the bottom of the tip, the active aggregation of cells to form tight cell-cell interactions drove the formation of spheroids. Using common laboratory micropipettes and pipette tips, the rate of spheroid generation and the generation reproducibility was characterized from five boxes each with 80 tips. The ease of transferring reagents allowed modeling of the growth of microvascular endothelial cells in tumor spheroids. Moreover, the pairing and fusion of tumor spheroids could be manipulated in the pipette tips, suggesting the potential for building and assembling heterogeneous micro-tumor tissues in vitro to mimic solid tumors in vivo. This study demonstrated that spheroids can be readily and cost-effectively generated in standard biological laboratories in a timely manner using pipette tips.

使用移液器吸头培养三维肿瘤球

作者:潘茸1,杨晓艳1,武士铭1,谢媛媛1,陈凤1,宁珂1,孙伟2,余玲1
机构:1西南大学,材料与能源学院,清洁能源与先进材料研究所,发光分析与分子传感教育部重点实验室,中国重庆,400715;2海南师范大学,化学化工学院,中国海口,571158
目的:三维细胞培养相较于常规的二维培养在模拟肿瘤微环境上具备很大的优势。本文旨在将移液器吸头(商品化且实验室通用的一种耗材)作为独特的细胞培养容器进行细胞培养,以实现高通量、简单、省时且经济高效的三维细胞球培养。
创新点:1.将移液器吸头作为细胞培养容器,其实验过程未涉及任何机械加工和化学处理,因此极大地简化了肿瘤球培养实验过程,为实验条件有限的实验室提供了三维细胞培养的替代方案;2.该培养平台集稳定、便捷、省时、低成本和高通量于一体;3.该平台在培养过程中可进行原位观测;4.除了用于细胞球的培养外,该培养方式还展示出在肿瘤球融合和药物筛选等方面的应用潜力。
方法:1.将细胞悬液吸到移液器吸头中,并将吸头置于吸头盒内,然后放入细胞培养箱里进行常规细胞培养。2.培养24 h后,在显微镜下观察细胞团聚形成三维细胞球;可直接推动移液器按钮进行细胞球的转移和后续分析。3.将两个或多个在吸头内形成的细胞球转移到同一个吸头,实现多个肿瘤球的配对和融合。4.待细胞成球后,在移液器吸头内加入药物,评价药物的细胞毒性。
结论:1.将移液器吸头作为三维细胞培养平台,通过简单地抽吸和培养,即可在短时间内高通量地获得细胞球。2.成功构建了两种肿瘤细胞(DU145)与血管内皮细胞(HUVEC)的共培养模型。3.两个或多个细胞球可通过配对和融合组装成更大的细胞聚集体,显示了该平台在微组织工程领域中的应用前景。4.成功在移液器吸头内完成了原位药物筛选;不同浓度的阿霉素会对移液器吸头中的细胞球造成不同程度的损伤。

关键词:移液器吸头;三维细胞培养;肿瘤球;共培养;原位观测

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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