Abstract
Mesenchymal stem cells (MSCs) have been widely studied with regard to their potential use in cell therapy protocols and regenerative medicine. However, a better comprehension about the factors and molecular mechanisms driving cell differentiation is now mandatory to improve our chance to manipulate MSC behavior and to benefit future applications. In this work, we aimed to study gene regulatory networks at an early step of osteogenic differentiation. Therefore, we analyzed both the total mRNA and the mRNA fraction associated with polysomes on human adipose tissue-derived stem cells (hASCs) at 24 h of osteogenesis induction. The RNA-seq results evidenced that hASC fate is not compromised with osteogenesis at this time and that 21 days of continuous cell culture stimuli are necessary for full osteogenic differentiation of hASCs. Furthermore, early stages of osteogenesis induction involved gene regulation that was linked to the management of cell behavior in culture, such as the control of cell adhesion and proliferation. In conclusion, although discrete initial gene regulation related to osteogenesis occur, the first 24 h of induction is not sufficient to trigger and drive in vitro osteogenic differentiation of hASCs.
Highlights
IntroductionMesenchymal stem cells (MSCs), including human adipose tissue-derived stem cells (hASCs), are undifferentiated cell populations characterized by the ability to undergo self-renewal and the capacity for multilineage differentiation1. hASCs can be found in adipose tissue in large amounts (millions to billions of cells), and they can be collected and harvested by a minimally invasive procedure
Mesenchymal stem cells (MSCs), including human adipose tissue-derived stem cells, are undifferentiated cell populations characterized by the ability to undergo self-renewal and the capacity for multilineage differentiation1. hASCs can be found in adipose tissue in large amounts, and they can be collected and harvested by a minimally invasive procedure
In a model of adipogenic differentiation, RNA sequencing (RNA-seq) analysis of the mRNA fraction associated with polysomes showed a significant percentage of regulated mRNAs that were controlled at the translational level and/or by changes in their transcript levels[12]
Summary
Mesenchymal stem cells (MSCs), including human adipose tissue-derived stem cells (hASCs), are undifferentiated cell populations characterized by the ability to undergo self-renewal and the capacity for multilineage differentiation1. hASCs can be found in adipose tissue in large amounts (millions to billions of cells), and they can be collected and harvested by a minimally invasive procedure. Little is known about how cell fate determinants are regulated in functionally important gene regulatory networks, which remains a challenge Gene expression analysis, both genome-wide and targeted at specific gene subsets, has played a key role in improving our understanding into the molecular pathways involved in hASCs self-renewal and differentiation[5]. In a model of adipogenic differentiation, RNA-seq analysis of the mRNA fraction associated with polysomes showed a significant percentage of regulated mRNAs that were controlled at the translational level and/or by changes in their transcript levels[12] This previous work demonstrated that three days of in vitro cell differentiation induction with adipogenic stimulation medium are sufficient for the initiation of adipogenesis and the upregulation of its correlated gene networks
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