posted on 2021-09-08, 18:38authored byDongguo Lin, Xiao Chen, Yang Liu, Zhun Lin, Yanzhang Luo, Mingpeng Fu, Na Yang, Dayu Liu, Jie Cao
Cancer stem cells (CSCs) are rare
and lack definite biomarkers,
necessitating new methods for a robust expansion. Here, we developed
a microfluidic single-cell culture (SCC) approach for expanding and
recovering colorectal CSCs from both cell lines and tumor tissues.
By incorporating alginate hydrogels with droplet microfluidics, a
high-density microgel array can be formed on a microfluidic chip that
allows for single-cell encapsulation and nonadhesive culture. The
SCC approach takes advantage of the self-renewal property of stem
cells, as only the CSCs can survive in the SCC and form tumorspheres.
Consecutive imaging confirmed the formation of single-cell-derived
tumorspheres, mainly from a population of small-sized cells. Through
on-chip decapsulation of the alginate microgel, ∼6000 live
cells can be recovered in a single run, which is sufficient for most
biological assays. The recovered cells were verified to have the genetic
and phenotypic characteristics of CSCs. Furthermore, multiple CSC-specific
targets were identified by comparing the transcriptomics of the CSCs
with the primary cancer cells. To summarize, the microgel SCC array
offers a label-free approach to obtain sufficient quantities of CSCs
and thus is potentially useful for understanding cancer biology and
developing personalized CSC-targeting therapies.