Microfluidic Device for Aptamer-Based Cancer Cell Capture and Genetic Mutation Detection.

Area of Science:

• Biotechnology
• Molecular Biology
• Genomics

Background:

• Cancer cell heterogeneity poses challenges for accurate genetic mutation analysis.
• Efficient isolation and amplification of genomic DNA (gDNA) from limited cell populations are crucial for cancer research.
• Existing methods may require substantial sample volumes, limiting analysis of rare cell populations.

Purpose of the Study:

• To develop and validate a microfluidic device for specific capture and in situ isolation of cancer cell genomic DNA (gDNA).
• To enable sensitive amplification and sequencing of target genes from isolated gDNA for mutation detection.
• To assess the utility of this platform for identifying TP53 gene mutations in cervical and ovarian cancer cells.

Main Methods:

• Immobilization of nucleic acid aptamers on micropillars for specific cancer cell capture.
• In situ cell lysis and gDNA isolation via physical entanglement in a secondary micropillar array.
• Selective gene amplification using multiple displacement amplification (MDA).
• Sequencing of amplified gDNA and comparison with wild-type sequences for mutation identification.

Main Results:

• Demonstrated successful capture and isolation of cancer cells and their gDNA within the microfluidic device.
• Validated the ability to amplify and sequence specific genes from the isolated gDNA.
• Identified TP53 gene mutations in tested cervical and ovarian cancer cell lines.
• Showcased the potential for monitoring multiple genetic mutations in small cell populations.

Conclusions:

• The developed microfluidic device provides an efficient method for cancer cell capture and gDNA isolation.
• This platform facilitates sensitive genetic mutation analysis from limited patient samples.
• The technology holds promise for personalized cancer diagnostics and monitoring due to its ability to detect diverse mutations.