Molecular cytogenetic characterization of the human cerebral microvessel endothelial cell line hCMEC/D3

Area of Science:

• Cell Biology
• Genetics
• Neuroscience

Background:

• The hCMEC/D3 cell line is a widely used model for the human blood-brain barrier (BBB).
• This cell line, typically used between passages 25 and 35, was believed to maintain a normal diploid karyotype.
• hCMEC/D3 cells retain many characteristics of primary human brain endothelial cells.

Purpose of the Study:

• To investigate the chromosomal integrity of the hCMEC/D3 cell line.
• To characterize the karyotype of hCMEC/D3 cells at passage 30 using advanced cytogenetic techniques.

Main Methods:

• High-resolution multicolor fluorescence in situ hybridization (FISH) was employed.
• Detailed analysis of numerical and structural chromosomal aberrations was performed.

Main Results:

• A complex karyotype was identified in hCMEC/D3 cells at passage 30.
• Clonal cryptic unbalanced structural rearrangements and numerical aberrations were detected.
• Specific chromosomal abnormalities include del(5)(q11), del(9)(q11), +9, del(11)(q13-14), der(14)t(14;21), der(15)t(9;15), dup(15), der(21)t(17;21), and -22.

Conclusions:

• The hCMEC/D3 cell line possesses a complex karyotype with clonal chromosomal rearrangements.
• The findings underscore the importance of molecular cytogenetics for validating cell lines before their use in critical research.
• Routine cytogenetic testing is recommended for all cell lines to ensure data reliability.