Cardiac differentiation of human induced Pluripotent Stem Cells in Vertical-Wheel bioreactors

Inês Alexandra Brites Ribeiro

Key information

Authors:

Inês Alexandra Brites Ribeiro (Inês Alexandra Brites Ribeiro)

Supervisors:

Carlos André Vitorino Rodrigues (Carlos André Vitorino Rodrigues)

Published in

12/06/2021

Abstract

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have great potential for applications in regenerative medicine, disease modeling and drug development, since they can be generated in a short time and in large quantities. Vertical-Wheel Bioreactors (VWBRs) have a gentle agitation system, being promising for the large-scale production of shear-stress sensitive cells, unlike the commonly used stirred tank bioreactors (STBRs). An economic model for the production of hiPSC-CMs in 3L VWBRs allowed to identify the reagents and the maturation phase as the main cost drivers. The minimum working volume of the bioreactor was found to be a limitation for some autologous processes, because of increased costs related to an excessive production of hiPSC-CMs arising from using a greater volume than necessary. Further studies conducted in VWBRs are necessary to assess the application of lower cell densities, together with the optimization of other parameters (e.g agitation speed), to avoid excess production costs while not compromising differentiation efficiency. Allogeneic therapies cost/dose were found to be lower than for autologous therapies at different scales. Additionally, hiPSCs were cultured in lab with three different culture media (mTesR1, E8 or B8) and differentiated into CMs via modulation of the canonical Wnt pathway in two distinct culture media (RPMI/B27 or CDM3). All conditions generated far lower percentages of CMs (up to 32.60% cTnT-positive cells) than the 80-98% cTnT-positive cells obtained in the original protocol after 15 days of differentiation, which might be associated with the lack of cell growth and adhesion to Matrigel substrate.