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Article

A Comparative Study of Cell Culture Conditions during Conversion from Primed to Naive Human Pluripotent Stem Cells

by
Irene Romayor
1,2,3,
Lara Herrera
1,2,
Maria Burón
1,2,
Myriam Martin-Inaraja
1,2,
Laura Prieto
1,2,
Jone Etxaniz
1,2,
Marta Inglés-Ferrándiz
1,2,
Jose Ramon Pineda
3,4 and
Cristina Eguizabal
1,2,*
1
Cell Therapy, Stem Cells and Tissues Group, Biocruces Bizkaia Health Research Institute, 48903 Barakaldo, Spain
2
Research Unit, Basque Centre for Blood Transfusion and Human Tissues, 48960 Galdakao, Spain
3
Cell Biology and Histology Department, University of the Basque Country (UPV/EHU), 48940 Leioa, Spain
4
Achucarro Basque Center for Neuroscience, University of the Basque Country (UPV/EHU), 48940 Leioa, Spain
*
Author to whom correspondence should be addressed.
Biomedicines 2022, 10(6), 1358; https://doi.org/10.3390/biomedicines10061358
Submission received: 12 May 2022 / Revised: 1 June 2022 / Accepted: 3 June 2022 / Published: 9 June 2022
(This article belongs to the Special Issue The Promise of Induced Pluripotent Stem Cells in the Biomedical Research)
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Abstract

The successful reprogramming of human somatic cells into induced pluripotent stem cells (hiPSCs) represented a turning point in the stem cell research field, owing to their ability to differentiate into any cell type with fewer ethical issues than human embryonic stem cells (hESCs). In mice, PSCs are thought to exist in a naive state, the cell culture equivalent of the immature pre-implantation embryo, whereas in humans, PSCs are in a primed state, which is a more committed pluripotent state than a naive state. Recent studies have focused on capturing a similar cell stage in human cells. Given their earlier developmental stage and therefore lack of cell-of-origin epigenetic memory, these cells would be better candidates for further re-differentiation, use in disease modeling, regenerative medicine and drug discovery. In this study, we used primed hiPSCs and hESCs to evaluate the successful establishment and maintenance of a naive cell stage using three different naive-conversion media, both in the feeder and feeder-free cells conditions. In addition, we compared the directed differentiation capacity of primed and naive cells into the three germ layers and characterized these different cell stages with commonly used pluripotent and lineage-specific markers. Our results show that, in general, naive culture NHSM medium (in both feeder and feeder-free systems) confers greater hiPSCs and hESCs viability and the highest naive pluripotency markers expression. This medium also allows better cell differentiation cells toward endoderm and mesoderm.
Keywords: hiPSCs; hESCs; pluripotency; naive status; primed status; cell differentiation; ectoderm; mesoderm; endoderm hiPSCs; hESCs; pluripotency; naive status; primed status; cell differentiation; ectoderm; mesoderm; endoderm

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MDPI and ACS Style

Romayor, I.; Herrera, L.; Burón, M.; Martin-Inaraja, M.; Prieto, L.; Etxaniz, J.; Inglés-Ferrándiz, M.; Pineda, J.R.; Eguizabal, C. A Comparative Study of Cell Culture Conditions during Conversion from Primed to Naive Human Pluripotent Stem Cells. Biomedicines 2022, 10, 1358. https://doi.org/10.3390/biomedicines10061358

AMA Style

Romayor I, Herrera L, Burón M, Martin-Inaraja M, Prieto L, Etxaniz J, Inglés-Ferrándiz M, Pineda JR, Eguizabal C. A Comparative Study of Cell Culture Conditions during Conversion from Primed to Naive Human Pluripotent Stem Cells. Biomedicines. 2022; 10(6):1358. https://doi.org/10.3390/biomedicines10061358

Chicago/Turabian Style

Romayor, Irene, Lara Herrera, Maria Burón, Myriam Martin-Inaraja, Laura Prieto, Jone Etxaniz, Marta Inglés-Ferrándiz, Jose Ramon Pineda, and Cristina Eguizabal. 2022. "A Comparative Study of Cell Culture Conditions during Conversion from Primed to Naive Human Pluripotent Stem Cells" Biomedicines 10, no. 6: 1358. https://doi.org/10.3390/biomedicines10061358

APA Style

Romayor, I., Herrera, L., Burón, M., Martin-Inaraja, M., Prieto, L., Etxaniz, J., Inglés-Ferrándiz, M., Pineda, J. R., & Eguizabal, C. (2022). A Comparative Study of Cell Culture Conditions during Conversion from Primed to Naive Human Pluripotent Stem Cells. Biomedicines, 10(6), 1358. https://doi.org/10.3390/biomedicines10061358

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