A Comparative Study of Cell Culture Conditions during Conversion from Primed to Naive Human Pluripotent Stem Cells.
Irene RomayorLara HerreraMaria BurónMyriam Martin-InarajaLaura PrietoJone EtxanizMarta Inglés-FerrándizJose Ramon PinedaCristina EguizabalPublished in: Biomedicines (2022)
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.
Keyphrases
- induced apoptosis
- pluripotent stem cells
- induced pluripotent stem cells
- hiv infected
- cell cycle arrest
- stem cells
- single cell
- endothelial cells
- cell therapy
- embryonic stem cells
- drug discovery
- dna methylation
- gene expression
- oxidative stress
- cell death
- poor prognosis
- mesenchymal stem cells
- working memory
- decision making
- cell proliferation
- long non coding rna
- binding protein