A Quality Assurance Scheme for Prenatal Detection of Aneuploidy Based on Developing Chimeric Quality-Control Cells

Area of Science:

• Genetics
• Molecular Biology
• Cell Biology

Background:

• Quality control for prenatal molecular diagnosis is limited by the scarcity of non-renewable rare disease samples.
• Aneuploid cell lines are needed for developing reliable quality control materials for prenatal aneuploidy detection.

Purpose of the Study:

• To prepare aneuploid amniocyte cell lines for use as quality control materials.
• To establish quality control cells for fluorescence in situ hybridization (FISH)-based aneuploidy detection.

Main Methods:

• Aneuploid amniotic fluid cells (47,XY,+18) were transfected with SV40 large T antigen gene vectors to create immortalized cell lines.
• Chimeric quality control cells were prepared by mixing immortalized aneuploid cells with normal amniocytes (46,XY) at various theoretical ratios.
• The feasibility of these chimeric cells as internal quality controls (IQC) for FISH detection was evaluated.

Main Results:

• Immortalized aneuploid amniocytes (SV40LT) allowed for indefinite culturing and preparation of quality control cells.
• FISH analysis confirmed actual ratios of aneuploid cells in chimeric samples were approximately 1.5%, 10.3%, 19.9%, and 40.8%.
• Fluorescence signals for key chromosomes (13, 18, 21, X, Y) in the chimeric cells were consistent with primary cells.

Conclusions:

• The developed aneuploid cell lines can potentially overcome the shortage of quality control materials for prenatal aneuploidy detection.
• This work provides a foundation for IQC-based detection methods in FISH analysis for chromosomal abnormalities.