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On-line Access: 2023-10-18

Received: 2022-11-27

Revision Accepted: 2023-03-28

Crosschecked: 2023-10-19

Cited: 0

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Citations:  Bibtex RefMan EndNote GB/T7714


Ling YU


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


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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journal="Journal of Zhejiang University Science A",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Using pipette tips to readily generate spheroids comprising single or multiple cell types
%A Rong PAN
%A Xiaoyan YANG
%A Shiming WU
%A Yuanyuan XIE
%A Feng CHEN
%A Wei SUN
%A Ling YU
%J Journal of Zhejiang University SCIENCE A
%V 24
%N 10
%P 875-885
%@ 1673-565X
%D 2023
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.A22D0235

T1 - Using pipette tips to readily generate spheroids comprising single or multiple cell types
A1 - Rong PAN
A1 - Xiaoyan YANG
A1 - Shiming WU
A1 - Yuanyuan XIE
A1 - Feng CHEN
A1 - Ke NING
A1 - Wei SUN
A1 - Ling YU
J0 - Journal of Zhejiang University Science A
VL - 24
IS - 10
SP - 875
EP - 885
%@ 1673-565X
Y1 - 2023
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.A22D0235

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.


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。


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


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