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A microfluidic platform enables comprehensive gene expression profiling of mouse retinal stem cells
Brenda L K Coles1, Mahmoud Labib2, Mahla Poudineh3
1Department of Molecular Genetics, University of Toronto, 1 King's College Circle, Toronto, ON M5S 1A8, Canada. derek.van.der.kooy@utoronto.ca.
Lab on a Chip
|October 15, 2021
Summary
Researchers developed a new microfluidic platform and cell-surface marker strategy to enrich rare retinal stem cells (RSCs). This breakthrough offers a promising method for cell replacement therapy in treating blindness caused by retinal degeneration.
Area of Science:
- Ophthalmology
- Regenerative Medicine
- Biotechnology
Background:
- Retinal degeneration leads to vision loss, with cell replacement therapy offering a potential solution.
- Identifying and isolating rare retinal stem cells (RSCs) for therapy is a significant challenge.
- Current methods lack validated markers for prospective enrichment of RSCs.
Purpose of the Study:
- To develop an efficient method for enriching retinal stem cells (RSCs).
- To identify novel cell-surface markers for RSC isolation.
- To characterize the molecular and functional properties of RSCs.
Main Methods:
- Development of a microfluidic platform integrating nickel micromagnets and herringbone structures.
- Implementation of an affinity enrichment strategy using cell-surface markers.
- Single-cell whole-transcriptome profiling to identify differentially expressed markers.
Main Results:
- The microfluidic platform achieved highly efficient enrichment of RSCs.
- Simultaneous targeting of three cell-surface markers enriched RSCs 1:3.
- Identification of four novel cell-surface markers yielded a 1:20 enrichment of RSCs.
- Discovered transcription factors crucial for RSC maintenance, quiescence, and proliferation.
Conclusions:
- The developed microfluidic and affinity-based strategy significantly enhances RSC enrichment.
- This approach provides a critical step towards clinical application of RSC-based therapies.
- The study offers a comprehensive molecular and functional profile of RSCs.
