Online monitoring of hiPSC expansion and hepatic differentiation in 3D culture by dielectric spectroscopy.
Inês A IsidroPedro VicenteDaniel A M PaisJoana I AlmeidaMara DominguesBernardo AbecasisNatalia Zapata-LinaresJuan Roberto Rodriguez-MadozFelipe ProsperAnders AspegrenPaula M AlvesMargarida SerraPublished in: Biotechnology and bioengineering (2021)
Hepatocyte-like cells derived from human-induced pluripotent stem cells (hiPSC-HLC) are expected to have important applications in drug screening and regenerative medicine. However, hiPSC-HLC are difficult to produce on a large-scale to obtain relevant numbers for such applications. The aim of this study was to implement a novel integrated strategy for scalable production of hiPSC-HLC and demonstrate the applicability of dielectric spectroscopy to monitor hiPSC expansion/differentiation processes. We cultured hiPSC as three-dimensional (3D) aggregates in stirred-tank bioreactors (STB) operated in perfusion with an in situ capacitance probe. Dissolved oxygen concentration and dilution rate were controlled along the process and after 5 days of cell expansion, the hepatic differentiation was integrated in sequential steps for 28 days. The hiPSC were able to grow as 3D aggregates and the expression of hepatic markers and albumin production after differentiation confirmed that hepatocyte differentiation improved when compared to 2D culture. These hiPSC-HLC exhibited functional characteristics of hepatocytes including glycogen storage and drug metabolization capacity. Our results also show a good correlation between the cell permittivity measured online and the aggregate biovolume measured by standard offline methods, demonstrating for the first time the potential of dielectric spectroscopy to monitor hiPSC expansion and differentiation in STB.
Keyphrases
- induced pluripotent stem cells
- high resolution
- single cell
- endothelial cells
- single molecule
- poor prognosis
- health information
- liver injury
- social media
- magnetic resonance imaging
- magnetic resonance
- drug induced
- stem cells
- computed tomography
- quantum dots
- wastewater treatment
- ms ms
- mesenchymal stem cells
- living cells
- liquid chromatography
- adverse drug
- human health
- fluorescent probe
- pluripotent stem cells