DAPT inhibits the chondrogenesis of human umbilical cord mesenchymal stem cells

Abstract

Notch signaling plays a key role in cell proliferation and differentiation, and is important in several biological processes, but its role in the chondrogenesis of human umbilical cord mesenchymal stem cells (UC-MSCs) is still unknown. N-[N-(3,5-difluorophenacetyl-L-alanyl)]-(S)-phenylglycinet-butyl ester (DAPT) is the inhibitor of Notch pathway. The aim of this study is to investgate the influence of DAPT on the chondrogenesis of UC-MSCs. In our study, UC-MSCs were isolated from human umbilical cord and their characteristics were identified. The UC-MSCs were induced to differentiate into chondrocytes in vitro and treated with 5 μM DAPT. Glycosaminoglycan (GAG) and collagen type II (COL-2A1) were analyzed qualitatively and quantitatively. The gene expression of Notch-1, Hes-1, GAG and COL-2A1 were analyzed by quantitative polymerase chain reaction (qPCR). The UC-MSCs separated from human umbilical cord, followed the characteristics of Mesenchymal Stem Cells (MSCs). The gene expression of Notch-1 and Hes-1 decreased after chondrogenic induction but the percentage in G1 period and the content of GAG and COL-2A1 increased. The expression of all tested Notch signaling and proliferation genes declined when 5 μM DAPT was added, also the content of GAG and COL-2A1 also decreased. Our study revealed that Notch signaling exists in UC-MSCs and it may remain the proliferative activity of UC-MSCs. Once the chondrogenesis begins, Notch signaling strength decline evidently. DAPT inhibits the chondrogenesis of UC-MSCs.

Cellular preconditioning and mesenchymal stem cell ferroptosis

“…The research interest in ferroptosis is gaining momentum as it is implicated in the poor engraftment of donor mesenchymal stem cells (MSCs), leading to the reduced efficiency of MSC-based treatment[ 6 , 10 ]. Ferroptosis can occur in response to diverse stimuli, such as starvation, exposure to toxins, ROS, etc , leading to intracellular iron accumulation[ 11 ].…”

“…However, recent research has shed light on some of the critical signaling pathways, such as p53, nuclear factor erythroid 2-related factor 2 (Nrf2), protein kinase R-like endoplasmic reticulum kinase[ 12 ], phospholipid remodeling pathway, GSH peroxidase 4 (GPX4), ferroptosis suppressor protein 1 and other emerging signaling pathways are considered to play a role therein[ 13 , 14 ]. This editorial aims to shed light on the pivotal role of the novel cystathionine γ-lyase (CSE)/hydrogen sulfide (H 2 S) pathway in ferroptosis, supported by the recent scientific evidence by Hu et al [ 10 ].…”

“…Bioinformatic studies revealed multiple genes expressed in ferroptosis and other genes, and microRNAs can regulate ferroptosis and disease progression[ 30 , 31 ]. Given the significant role of the CSE/H 2 S pathway in cellular function, recent studies have revealed its crucial role in regulating ferroptosis in stem cells[ 10 , 32 ].…”

“…In their comprehensive investigation, Hu et al [ 10 ] investigated the therapeutic potential of human umbilical cord MSCs (hUCMSCs) in treating hypoxia-induced pulmonary arterial hypertension (PAH) in a murine model. They explored the impact of modulating the CSE/H 2 S pathway in preventing ferroptosis in hUCMSCs.…”

“…Remarkably, preconditioning hUCMSCs with a ferroptosis inhibitor, Fer-1, exhibited enhanced survival and efficacy in managing PAH, while ferroptosis induced low cellular viability and therapeutic effectiveness. These compelling findings imply a potential association between ferroptosis and the survival of hUCMSCs in hypoxia-induced PAH[ 10 ].…”