Facile and Rapid Generation of Large-Scale Microcollagen
Gel Array for Long-Term Single-Cell 3D Culture and Cell Proliferation
Heterogeneity Analysis
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Abstract
Microfabricated devices are suitable
for single-cell analysis due
to their high throughput, compatible dimensions and controllable microenvironment.
However, existing devices for single-cell culture and analysis encounter
some limitations, such as nutrient depletion, random cell migration
and complicated fluid shear influence. Moreover, most of the single-cell
culture and analysis devices are based on 2D cell culture conditions,
even though 3D cell culture methods have been demonstrated to better
mimic the real cell microenvironment in vivo. To solve these problems,
herein we develop a microcollagen gel array (μCGA) based approach
for high-throughput long-term single-cell culture and single-cell
analysis under 3D culture conditions. Type-I collagen, a well-established
3D cell culture medium, was used as the scaffold for 3D cell growth.
A 2 × 2 cm PDMS chip with 10 000 μCGA units was
fabricated to encapsulate thousands of single cells in less than 15
min. Single cells were able to be confined and survive in μCGA
units for more than 1 month. The capability of large-scale and long-term
single-cell 3D culture under open culture conditions allows us to
study cellular proliferation heterogeneity and drug cytotoxicity at
the single-cell level. Compared with existing devices for single-cell
analysis, μCGA solves the problems of nutrient depletion and
random cellular migration, avoids the influence of complicated fluid
shear, and mimics the real 3D growth environment in vivo, thereby
providing a feasible 3D long-term single-cell culture method for single-cell
analysis and drug screening