Altered gene expression in lymphoblastoid cell lines after subculture

Area of Science:

• Cell Biology
• Genomics
• Immunology

Background:

• Lymphoblastoid cell lines (LCLs) are widely used for gene expression studies due to their immortalized nature.
• However, the stability of LCL cellular features, particularly gene expression, over time in culture is not well-characterized.

Purpose of the Study:

• To investigate the stability of gene expression in LCLs across different passage numbers.
• To evaluate the suitability of commonly used housekeeping genes, such as GAPDH, as internal controls in LCL gene expression analyses.

Main Methods:

• Gene expression analysis of LCLs at various passage numbers.
• Quantitative assessment of specific gene expression levels, including glyceraldehyde-3-phosphate dehydrogenase (GAPDH).

Main Results:

• Significant alterations in gene expression profiles were observed within the first 10 passages of LCL subculture.
• Expression levels of GAPDH, a commonly used housekeeping gene, demonstrated considerable variability during subculturing.
• These findings indicate that GAPDH may not be a reliable internal control for gene expression studies in LCLs.

Conclusions:

• Gene expression in LCLs is not stable and changes substantially with increasing passage numbers.
• The variability in GAPDH expression suggests caution is needed when using it as an internal control in LCL studies.
• Researchers should carefully validate housekeeping genes for stability in LCLs at specific passage ranges for accurate gene expression analysis.