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A Simple Red Blood Cell Lysis Method for the Establishment of B Lymphoblastoid Cell Lines
Published on: January 14, 2017
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Robust imaging and gene delivery to study human lymphoblastoid cell lines
Lachlan A Jolly1,2, Ying Sun3,4, Renée Carroll3,4
1School of Medicine, The University of Adelaide, Adelaide, 5005, Australia. Lachlan.Jolly@adelaide.edu.au.
Journal of Human Genetics
|June 22, 2018
Summary
We optimized methods for using lymphoblastoid cell lines (LCLs) in functional genomics. New techniques allow LCLs to adhere for sub-cellular studies and improve gene delivery efficiency for mechanistic research.
Area of Science:
- Cell Biology
- Genomics
- Molecular Biology
Background:
- Lymphoblastoid cell lines (LCLs) are widely used for genetic variation studies.
- Limited use of LCLs in functional genomics due to technical challenges.
- Challenges include non-adherent nature and poor gene transfection efficiency.
Purpose of the Study:
- To develop and optimize methods for LCLs in functional genomics.
- To enable sub-cellular spatial information analysis in LCLs.
- To improve gene delivery and cell purification for LCLs.
Main Methods:
- Developed a robust method for adhering LCLs to coverslips.
- Optimized lentiviral gene delivery to LCLs through extensive testing.
- Established cell purification strategies for high-purity LCL cultures.
Main Results:
- Achieved improved LCL adherence, preserving cellular integrity and morphology.
- Increased lentiviral transduction efficiency in LCLs from 3% to 48%.
- Developed methods for sustainable cultures with >85% transduced cells.
Conclusions:
- Provided optimized methodologies for LCLs in functional cell and molecular biology.
- These methods facilitate characterization of genetic variants and disease pathways.
- Enables high-throughput discovery of genetic modifiers in disease states.
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