# Using Microfluidic Single Cell Culture to Characterize Cancer Cell Asymmetric Division

> **NIH NIH R01** · MAGEE-WOMEN'S RES INST AND FOUNDATION · 2020 · $357,769

## Abstract

ABSTRACT
Whether cancer follows a hierarchical (linear) or stochastic (random) model of differentiation has critical clinical
implications, yet remains highly controversial. The hierarchical model mirrors normal stem cell biology with
cancer stem-like cells (CSC) following an established pattern of asymmetric divisions to give rise to distinct
progeny. In this model only CSC can initiate recurrent disease. In the stochastic model, any given tumor cell
can asymmetrically divide to give rise to the other cells of the tumor and initiate recurrent disease. The key to
determining the pattern of differentiation is characterizing asymmetric division potential. However the
asymmetric division potential of primary human cancer cells has not been directly interrogated due to a lack of
technology. We hypothesize that cancer cells will follow a hierarchical differentiation pathway, but rare
stochastic dedifferentiation events occur allowing some cells to obtain a CSC state. We further hypothesize
that factors regulating the asymmetric division of cancer cells, both hierarchical and stochastic, will significantly
impact cancer growth and represent therapeutic targets. To address the technology deficiency, we propose
SA1: To develop a high-throughput single-cell microfluidic culture device with selective live-cell
retrieval. A highly parallel, compact, 1024 microwell design will allow efficient single cell capture, growth, and
characterization of asymmetric division of primary human cancer cells. Automated data analysis and selective
single-cell retrieval capacity will allow for high-throughput functional and molecular analysis of common and
rare events. In order to validate this important new technology we will use ovarian cancer as a model system.
We propose: SA2: To directly interrogate the asymmetric division potential of primary ovarian cancer
cells. Our preliminary data indicates that ovarian cancer follows a hierarchical differentiation model with rare
(~1/3000) putative stochastic ‘dedifferentiation’ events wherein a progenitor cell gives rise to putative CSC. We
will expand our studies of the asymmetric division potential to assess inter-patient variability and determine the
impact of environmental stresses on dedifferentiation rates. Critically, using selective cell retrieval we can
evaluate the biologic characteristics (chemotherapy resistance, tumor initiation capacity) of single cells derived
following an asymmetric division to confirm the biologic implications of differentiation and dedifferentiation
events. Finally, we propose SA3: To identify regulators of cancer cell differentiation and drivers of
dedifferentiation. In parallel to the functional studies in SA2, mother and daughter cells from asymmetric
divisions will be isolated and expression of distinct stem cell related genes assessed via single cell qRT-PCR.
Factors differentially expressed between mother and daughter cells will then be evaluated for their impact on
CSC growth and differen...

## Key facts

- **NIH application ID:** 9851385
- **Project number:** 5R01CA203810-04
- **Recipient organization:** MAGEE-WOMEN'S RES INST AND FOUNDATION
- **Principal Investigator:** Ronald J Buckanovich
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $357,769
- **Award type:** 5
- **Project period:** 2017-02-01 → 2022-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9851385

## Citation

> US National Institutes of Health, RePORTER application 9851385, Using Microfluidic Single Cell Culture to Characterize Cancer Cell Asymmetric Division (5R01CA203810-04). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/9851385. Licensed CC0.

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