Scalable platform for human embryonic stem cell differentiation to cardiomyocytes in suspended microcarrier cultures

Marti Lecina, Sherwin Ting, Andre Choo, Shaul Reuveny, Steve Oh

Producción científica: Artículo en revista indizadaArtículorevisión exhaustiva

98 Citas (Scopus)

Resumen

A scalable platform for human embryonic stem cell (hESC)-derived cardiomyocyte (CM) production can provide a readily available source of CMs for cell therapy, drug screening, and cardiotoxicity tests. We have designed and optimized a scalable platform using microcarrier cultures in serum-free media supplemented with SB203580 mitogen-activated protein kinase-inhibitor. Different microcarriers (DE-53, Cytodex-1 and 3, FACT, and TOSOH-10) were used to investigate the effects of type, size, shape, and microcarrier concentrations on the differentiation efficiency. hESCs propagated on TOSOH-10 (protamine derivatized 10-μm beads) at the concentration of 0.125mg/mL produced 80% beating aggregates, threefold cell expansion, and 20% of CMs (determined by fluorescence-activated cell sorting for myosin heavy chain and α-actinin expression). The ratio of CM/hESC seeded in this system was 0.62 compared to 0.22 in the embryoid body control cultures. The platform robustness has been tested with HES-3 and H1 cell lines, and its scalability was demonstrated in suspended spinner cultures. However, spinner culture yields dropped to 0.33 CM/hESC probably due to shear stress causing some cell death. Cells dissociated from differentiated aggregates showed positive staining for cardio-specific markers such as α-actinin, myosin heavy and light chain, troponin I, desmin, and emilin-2. Finally, CM functionality was also shown by QT-prolongation (QTempo) assay with/without Astemizole. This study represents a new scalable bioprocessing system for CM production using reagents that can comply with Good Manufacturing Practice.

Idioma originalInglés
Páginas (desde-hasta)1609-1619
Número de páginas11
PublicaciónTissue Engineering - Part C: Methods
Volumen16
N.º6
DOI
EstadoPublicada - 1 dic 2010
Publicado de forma externa

Huella

Profundice en los temas de investigación de 'Scalable platform for human embryonic stem cell differentiation to cardiomyocytes in suspended microcarrier cultures'. En conjunto forman una huella única.

Citar esto