Forced differentiation in vitro leads to stress-induced activation of DNA damage response in hiPSC-derived chondrocyte-like cells.

Ewelina Stelcer,Tomasz Trzeciak,Magdalena Richter,Karol Jopek,Wiktoria Maria Suchorska,Marcin Rucinski,Katarzyna Kulcenty

doi:10.1371/journal.pone.0198079

Ewelina Stelcer, Tomasz Trzeciak + Show 5 more

Open Access

https://doi.org/10.1371/journal.pone.0198079

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Abstract

A human induced pluripotent stem cell line (GPCCi001-A) created by our group was differentiated towards chondrocyte-like cells (ChiPS) via monolayer culturing with growth factors. ChiPS are promising because they have the potential to be used in tissue engineering to regenerate articular cartilage. However, their safety must be confirmed before they can be routinely used in regenerative medicine. Using microarray analysis, we compared the ChiPS to both GPCCi001-A cells and chondrocytes. The analysis showed that, compared to both GPCCi001-A cells and chondrocytes, the expression of genes engaged in DNA damage and in the tumor protein p53 signalling pathways was significantly higher in the ChiPS. The significant amount of DNA double strand breaks and increased DNA damage response may lead to incomplete DNA repair and the accumulation of mutations and, ultimately, to genetic instability. These findings provide evidence indicating that the differentiation process in vitro places stress on human induced pluripotent stem cells (hiPSCs). The results of this study raise doubts about the use of stem cell-derived components given the negative effects of the differentiation process in vitro on hiPSCs.

Highlights

Stem cells (SCs), human induced pluripotent SCs, constitute a real hope to better understand the pathogenesis and improve the treatment of many disorders that are unresponsive to current treatments [1]
This study evaluated chondrocyte-like cells (ChiPS) derived from the GPCCi001-A cell line via a chondrogenic process
We found that the chondrogenic process in vitro is controlled by p53, which is mainly responsible for diminishing pluripotency factors and for preventing hiPSCderived cells to backslide into the pluripotency state

Summary

Introduction

Stem cells (SCs), human induced pluripotent SCs (hiPSCs), constitute a real hope to better understand the pathogenesis and improve the treatment of many disorders (e.g. neurodegenerative, neurovascular, and cardias diseases) that are unresponsive to current treatments [1]. HiPSCs hold great potential in regenerative medicine due to their potentially unlimited self-renewal capacity and ability to give rise to all of the somatic lineages in the body [2]. HiPSCs can be cultured in two main ways: feeder-dependent and feeder-free systems. In the feeder-dependent system, the cells are placed on a layer of inactivated murine embryonic. Differentiation leads to stress-induced activation of DDR in hiPSC-derived chondrocyte-like cells

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