The AC16 Cell Line Revisited: Integrated Cytogenetic, Genomic, and Transcriptomic Analysis Reveals a Highly Rearranged Near Tetraploid Genome

ABSTRACT: The human AC16 cell line was established in 2005 by fusing primary cells from adult human ventricular heart tissues with SV40 transformed, uridine auxotroph human fibroblasts devoid of mitochondrial DNA. This cell line is widely used as an in vitro model for studying cardiac biology, cardiotoxicity, and molecular mechanisms underlying cardiovascular diseases. However, despite its broad application, the detailed genetic and cytogenetic characteristics of this cell line has so far remained largely undefined. In the present study, we performed a comprehensive molecular and cytogenetic characterization of AC16 cells using multiple complementary approaches. Short tandem repeat profiling confirmed the AC16 reference profile deposited in the Cellosaurus database. Morphological examination by light and electron microscopy demonstrated a spindle-shaped, fibroblast-like morphology with a metabolically active cytoplasm containing numerous mitochondria, lysosomes, and extensive rough endoplasmic reticulum. Cytogenetic analyses using multicolor banding and inverted DAPI banding revealed an instable highly complex near-tetraploid karyotype with 90-100 chromosomes and numerous structural abnormalities, including deletions, derivative chromosomes, translocations, inverted duplications, and Robertsonian translocations. These findings were further supported by array comparative genomic hybridization, which demonstrated widespread genomic copy number alterations affecting nearly all chromosomes. This highlights that this cell line is rather suited as a cancer model than a model for normal heart cells. Next-generation sequencing confirmed the expression of several genes associated with cardiomyocyte function, including GJA1, GATA4, NFATC4, and CACNA1C, although classical cardiac markers such as TNNT2 and ACTN2 were absent or expressed at very low levels. Our findings reveal that AC16 cells possess a highly rearranged genome and exhibit a mixed molecular phenotype, emphasizing the importance of careful interpretation when using this cell line as a model for cardiac biology.

ORGANISM(S): Homo sapiens

PROVIDER: GSE335601 | GEO | 2026/06/30

REPOSITORIES: GEO