title: In vitro expansion of human cardiac progenitor cells: exploring 'omics tools for characterization of cell-based allogeneic products.
abstract: Human cardiac stem/progenitor cells (hCPCs) have been shown to be capable to regenerate contractile myocardium. However, because of their relative low abundance in the heart, in vitro expansion of hCPC is mandatory to achieve necessary quantities for allogeneic or autologous cardiac regeneration therapy applications (10(6)-10(9) cells/patient). Up to now, cell number requirements of ongoing phase I/IIa trials have been fulfilled with production in static monolayer cultures. However, this manufacturing process poses critical limitations when moving to the following clinical phases where hundreds of patients will be enrolled. For this, increased process yield is required, while guaranteeing the quality of the cell-based products. In this work, we developed and validated a robust, scalable, and good manufacturing practice (GMP)-compatible bioprocess for the expansion of high-quality hCPC. We applied platforms extensively used by the biopharmaceutical industry, such as microcarrier technology and stirred systems, and assessed culture conditions' impact on hCPC's quality and potency, as required by regulatory agencies. Complementary analytical assays including gene expression microarrays and mass spectrometry-based approaches were explored to compare transcriptome, proteome, surface markers, and secretion profiles of hCPC cultured in static monolayers and in stirred microcarrier-based systems. Our results show that stirred microcarrier-based culture systems enabled achieving more than 3-fold increase in hCPC expansion, when compared with traditional static monolayers, while retaining cell's phenotype and similar "omics" profiles. These findings demonstrate that this change in the production process does not affect cell's identity and quality, with potential to be translated into a transversal production platform for clinical development of stem-cell therapies.
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal; iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.
authors.forename: P
authors.initials: P
authors.lastname: Gomes-Alves
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal; iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.
authors.forename: M
authors.initials: M
authors.lastname: Serra
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal; iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.
authors.forename: C
authors.initials: C
authors.lastname: Brito
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal; iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.
authors.forename: C P
authors.initials: CP
authors.lastname: Ricardo
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal; iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.
authors.forename: R
authors.initials: R
authors.lastname: Cunha
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal; iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.
authors.forename: M F
authors.initials: MF
authors.lastname: Sousa
authors.affiliation: Coretherapix, Tres Cantos, Madrid, Spain.
authors.forename: B
authors.initials: B
authors.lastname: Sanchez
authors.affiliation: Centro Nacional de Biotecnología, Madrid, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Bernad
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal; Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Monte da Caparica, Portugal.
authors.forename: M J T
authors.initials: MJ
authors.lastname: Carrondo
authors.affiliation: Coretherapix, Tres Cantos, Madrid, Spain.
authors.forename: L
authors.initials: L
authors.lastname: Rodriguez-Borlado
authors.affiliation: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal; iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal. Electronic address: marques@ibet.pt.
authors.forename: P M
authors.initials: PM
authors.lastname: Alves
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D020869:Gene Expression Profiling; D006801:Humans; D013058:Mass Spectrometry; D008863:Microspheres; D009206:Myocardium / Q000201:enzymology*; D010641:Phenotype; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D013234:Stem Cells / Q000166:cytology*; D014184:Transplantation, Homologous
chemical_list: D015415:Biomarkers; D020543:Proteome