@article{3cd056d6c4b649b6a0082e9035d2cfae,
title = "Online monitoring of hiPSC expansion and hepatic differentiation in 3D culture by dielectric spectroscopy",
abstract = "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.",
keywords = "dielectric spectroscopy, human pluripotent stem cells-derived hepatocytes, integrated bioprocess, perfusion, stirred-tank bioreactor",
author = "Isidro, {In{\^e}s A.} and Pedro Vicente and Pais, {Daniel A.M.} and Almeida, {Joana I.} and Mara Domingues and Bernardo Abecasis and Natalia Zapata-Linares and Rodriguez-Madoz, {Juan R.} and Felipe Prosper and Anders Aspegren and Alves, {Paula M.} and Margarida Serra",
note = "Funding Information: The authors would like to acknowledge Eng Marcos Sousa for his technical assistance on bioreactor experiments as well as Francisca Arez, Dr. Sofia Rebelo, and Dr. Catarina Brito for their scientific support related to hepatic cell culture and characterization. We would like to thank Marc Torres and Jochen Uhlenk?ken at Hamilton for the support and useful discussions on the capacitance probe. We also acknowledge the Histopathology Facility at the Instituto Gulbenkian de Ci?ncia sample processing and imaging of H&E, PAS, and?MT?staining. This study was funded by a grant from ERA-NET E-Rare 3 research program, JTC ERAdicatPH (E-Rare3/0002/2015) and Funda??o para a Ci?ncia e Tecnologia project MetaCardio (PTDC/BTM-SAL/32566/2017); iNOVA4Health ? UIDB/04462/2020 and UIDP/04462/2020, a program financially supported by Funda??o para a Ci?ncia e Tecnologia/Minist?rio da Ci?ncia, Tecnologia e Ensino Superior, through national funds is acknowledged. P. V., J. I. A., and D. A. M. P. were supported by FCT fellowships SFRH/BD/145767/2019, SFRH/BD/116780/2016, and SFRH/BD/105873/2014, respectively. Funding Information: The authors would like to acknowledge Eng Marcos Sousa for his technical assistance on bioreactor experiments as well as Francisca Arez, Dr. Sofia Rebelo, and Dr. Catarina Brito for their scientific support related to hepatic cell culture and characterization. We would like to thank Marc Torres and Jochen Uhlenk{\"u}ken at Hamilton for the support and useful discussions on the capacitance probe. We also acknowledge the Histopathology Facility at the Instituto Gulbenkian de Ci{\^e}ncia sample processing and imaging of H&E, PAS, and MT staining. This study was funded by a grant from ERA‐NET E‐Rare 3 research program, JTC ERAdicatPH (E‐Rare3/0002/2015) and Funda{\c c}{\~a}o para a Ci{\^e}ncia e Tecnologia project MetaCardio (PTDC/BTM‐SAL/32566/2017); iNOVA4Health – UIDB/04462/2020 and UIDP/04462/2020, a program financially supported by Funda{\c c}{\~a}o para a Ci{\^e}ncia e Tecnologia/Minist{\'e}rio da Ci{\^e}ncia, Tecnologia e Ensino Superior, through national funds is acknowledged. P. V., J. I. A., and D. A. M. P. were supported by FCT fellowships SFRH/BD/145767/2019, SFRH/BD/116780/2016, and SFRH/BD/105873/2014, respectively. Publisher Copyright: {\textcopyright} 2021 Wiley Periodicals LLC",
year = "2021",
month = sep,
doi = "10.1002/bit.27751",
language = "English",
volume = "118",
pages = "3610--3617",
journal = "Biotechnology and Bioengineering",
issn = "0006-3592",
publisher = "Wiley-Blackwell",
number = "9",
}
