# Anchorage-independent culture for maintaining vocal fold epithelial stem cells

> **NIH NIH R21** · UNIVERSITY OF WISCONSIN-MADISON · 2020 · $191,250

## Abstract

Project Summary
Voice impairment (dysphonia) affects an estimated 20 million people in the United States and, despite
advances in diagnosis and treatment, remains a challenging clinical problem with a negative impact on patient
quality-of-life. Advances in the emerging field of VF mucosal biology have resulted in a number of powerful and
widely used in vitro experimental tools, such as human VF fibroblast cell lines and vibrational bioreactors;
however, there remain limited cell biology resources specific to the VF epithelium and its constituent cells,
hampering advances in VF epithelial biology and the understanding of epithelial diseases. A central reason for
this lack of progress is the technical challenges associated with VF epithelial cell isolation, maintenance, and
expansion in vitro. A reliable and reproducible culture methodology is needed to overcome these challenges. In
the proposed work, we will introduce and validate a promising in vitro technique – anchorage-independent
culture – for the isolation and maintenance of VF epithelial stem cells. Our team’s prior work in other epithelial
systems, and pilot data generated using VF epithelial cells, demonstrate that this technique results in the
formation of three-dimensional culture spheres that are enriched for tissue- specific stem cells. In the proposed
research, we will characterize the makeup, growth, maintenance, and passage of VF spheres, as well as test
the capacity of sphere-contained stem cells for in vitro differentiation and as a cell source for VF tissue
engineering. This work, conducted using human cells, will result in a validated and reproducible in vitro
methodology that will advance progress in VF epithelial (stem) cell biology and facilitate ongoing and future
research amongst the wider scientific community. This research effort represents a powerful collaboration
between experts in VF mucosal biology and stem cell biology and has broad applicability to both basic
scientific research and regenerative medicine of the larynx.

## Key facts

- **NIH application ID:** 9903281
- **Project number:** 5R21DC017836-02
- **Recipient organization:** UNIVERSITY OF WISCONSIN-MADISON
- **Principal Investigator:** Xudong Shi
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $191,250
- **Award type:** 5
- **Project period:** 2019-04-01 → 2022-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9903281

## Citation

> US National Institutes of Health, RePORTER application 9903281, Anchorage-independent culture for maintaining vocal fold epithelial stem cells (5R21DC017836-02). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/9903281. Licensed CC0.

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