An integrated microfluidics platform with high-throughput single-cell cloning array and concentration gradient generator for efficient cancer drug effect screening

Biao Wang; Bang-Shun He; Xiao-Lan Ruan; Jiang Zhu; Rui Hu; Jie Wang; Ying Li; Yun-Huang Yang; Mai-Li Liu

doi:10.1186/s40779-022-00409-9

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An integrated microfluidics platform with high-throughput single-cell cloning array and concentration gradient generator for efficient cancer drug effect screening

RESEARCH | Updated：2023-07-21

- An integrated microfluidics platform with high-throughput single-cell cloning array and concentration gradient generator for efficient cancer drug effect screening
- Military Medical Research Vol. 10, Issue 3, Pages: 325-341(2023)
- Affiliations：
  
  1.Key Laboratory of Magnetic Resonance in Biological Systems, State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, National Center for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Innovation Academy for Precision Measurement Science and Technology-Wuhan National Laboratory for Optoelectronics, Chinese Academy of Sciences, Wuhan 430071, China
  2.Department of Laboratory Medicine, Nanjing First Hospital, Nanjing Medical University, Nanjing 210006, China
  3.Department of Hematology, Renmin Hospital, Wuhan University, Wuhan 430060, China
  4.University of Chinese Academy of Sciences, Beijing 10049, China
  5.Canary Center at Stanford for Cancer Early Detection, Department of Radiology, School of Medicine, Stanford University, Palo Alto, CA 94304, USA
- Author bio：
  
  * bhe@njmu.edu.cn;
  liying@wipm.ac.cn
- Funds：
- DOI：10.1186/s40779-022-00409-9
  CLC：
- Published：2023-06
- Accepted：
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Cite this article as: Wang B, He BS, Ruan XL, Zhu J, Hu R, Wang J, et al. An integrated microfluidics platform with high-throughput single-cell cloning array and concentration gradient generator for efficient cancer drug effect screening. Mil Med Res. 2022;9(1):51.
DOI：

Cite this article as: Wang B, He BS, Ruan XL, Zhu J, Hu R, Wang J, et al. An integrated microfluidics platform with high-throughput single-cell cloning array and concentration gradient generator for efficient cancer drug effect screening. Mil Med Res. 2022;9(1):51. DOI： 10.1186/s40779-022-00409-9.

摘要

Abstract

Background:

Tumor cell heterogeneity mediated drug resistance has been recognized as the stumbling block of cancer treatment. Elucidating the cytotoxicity of anticancer drugs at single-cell level in a high-throughput way is thus of great value for developing precision therapy. However

current techniques suffer from limitations in dynamically characterizing the responses of thousands of single cells or cell clones presented to multiple drug conditions.

Methods:

We developed a new microfluidics-based "SMART" platform that is Simple to operate

able to generate a Massive single-cell array and Multiplex drug concentrations

capable of keeping cells Alive

Retainable and Trackable in the microchambers. These features are achieved by integrating a Microfluidic chamber Array (4320 units) and a six-Concentration gradient generator (MAC)

which enables highly efficient analysis of leukemia drug effects on single cells and cell clones in a high-throughput way.

Results:

A simple procedure produces 6 on-chip drug gradients to treat more than 3000 single cells or single-cell derived clones and thus allows an efficient and precise analysis of cell heterogeneity. The statistic results reveal that Imatinib (Ima) and Resveratrol (Res) combination treatment on single cells or clones is much more efficient than Ima or Res single drug treatment

indicated by the markedly reduced half maximal inhibitory concentration (IC

). Additionally

single-cell derived clones demonstrate a higher IC

in each drug treatment compared to single cells. Moreover

primary cells isolated from two leukemia patients are also found with apparent heterogeneity upon drug treatment on MAC.

Conclusions:

This microfluidics-based "SMART" platform allows high-throughput single-cell capture and culture

dynamic drug-gradient treatment and cell response monitoring

which represents a new approach to efficiently investigate anticancer drug effects and should benefit drug discovery for leukemia and other cancers.

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references

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