Pluripotent Cell Technology

The main function of the Pluripotent Cell Technology Unit (PCTUnit) is to provide technological innovation, support and scientific services to CNIC researchers in the generation of biological models, in vivo and/or in vitro, through the manipulation of mouse embryonic stem  cells (mESC) and human induced pluripotent cells (hiPSC).  With the purpose of ensuring an adequate workplace for researchers, the Unit supervises and manages the proper functioning of two culture rooms, one devoted to human stem cells and the other to mouse embryonic stem cells. The PCTUnit staff offer personalized training programs and advises, coordinate different and specific experimental trials, and provides validated and standardized reagents.

Since its formation, the Unit has contributed to the generation of numerous lines of genetically modified mice (knockout/in, conventional and conditional mice), through homologous recombination in mESC, which have been of significant support in many development and cardiovascular diseases research projects. Currently, the Unit also contributes to the generation of genetically modified mESC lines using CRISPR/Cas9 technology and assists the researchers in fine-tuning differentiation protocols for a specific lineage.

The CRISPR/Cas9-based gene editing tool together with somatic cell reprogramming offers a great opportunity in translational medicine, enabling the study of disease models in vitro.  In this area, the Unit assists the researchers in the derivation of hiPSC from dermal fibroblasts of patients affected by cardiovascular diseases. Then, by inserting mutations in a wildtype hiPSC line or by correcting mutations in patients derived hiPSC, we contribute to the generation of isogenic hiPSC and develop differentiation programs to cardiac lineage. The Unit offers comprehensive support in all experimental processes, including design, transfection, selection and genotyping of gene edited hiPSCs.

The genetic modification mediated by the CRISPR/Cas9 system is also being used to generate transgenic pig models. The Unit has recently joined its expertise with CNIC researchers in the development of projects based on such genetic models, extremely valuable in translational cardiovascular medicine.