Diamond-Blackfan Anemia (DBA) is a rare congenital bone marrow failure syndrome, associated with mutations in genes encoding ribosomal proteins, including R180C mutation in RPL27 gene. This mutation is present in less than 1% of cases; therefore, to investigate it, an in vitro model of the mutation was created
using embryonic stem cells (ESC).

For this purpose, two new cell lines were established: B9, carrying the RPL27_R180C mutation, and 1B11, wild-type control line. A series of experiments was conducted to verify the cells’ suitability for further research.

To confirm that pluripotency was maintained after genetic intervention, immunohistochemical staining was performed. The presence of the mutation in B9 cells and its absence in 1B11 were verified using genotyping. Along with that, cellular respiration was compared in mutant and wild-type cells using Seahorse XF Analyzer, in order to determine whether the energy-producing pathways are affected by the mutation.

The results demonstrate that ESC lines B9 and 1B11 meet the established standards for pluripotent cells according to literature protocols. Comparison of cellular respiration showed that mutant cells exhibit respiration rates comparable to wild-type cells and rely on similar energy-producing pathways. Within the framework of the research, two new ESC lines, B9 and 1B11, have been characterized and confirmed to be suitable for DBA disease modeling. This enables a closer view at the mutation and its effect on the cell function and may contribute to a deeper understanding of DBA pathogenesis and the development of improved therapeutic strategies.