Stable overexpression of p130/E2F4 affects the multipotential abilities of bone‐marrow‐derived mesenchymal stem cells

Abstract

Bone-marrow-derived mesenchymal stem cells (MSCs) have great potential in transplantation medicine due to their multiple advantages. However, the controlled differentiation of MSCs is one of the key aspects of effective clinical transplantation. Growing evidence suggests that the cell cycle plays an important role in regulating differentiation, while p130 and E2F4 are key to cell cycle checkpoints. The aim of the study is to evaluate the effects and mechanism of p130/E2F4 on the multidifferentiation of MSCs. Our data showed that the transduction efficiencies of p130 or E2F4 mediated by lentiviral vectors were 80.3%-84.4%. p130 and E2F4 mRNA expression was significantly higher in MSC-p130 and MSC-E2F4 cells than in MSC normal control (NC) cells. Similar results were also observed for p130 and E2F4 protein expression. After osteogenic or adipogenic differentiation, the G1 phase was significantly delayed in the MSC-p130 and MSC-E2F4 groups compared with that in the MSC-NC group. However, the G1 phase in the MSC-p130 and MSC-E2F4 groups did the opposite after chondrogenic differentiation. Moreover, overexpressing p130 or E2F4 significantly improved osteogenic differentiation while inhibiting adipogenic and chondrogenic differentiation of mouse MSCs (mMSCs). Moreover, overexpressing p130 or E2F4 significantly improved migration but not proliferation of mMSCs. Our data suggest that cell cycle regulation may be involved in p130/E2F4-mediated changes in the multipotential abilities of bone-marrow-derived mMSCs.

RETRACTED ARTICLE: Overexpression of TGFβ1 in murine mesenchymal stem cells improves lung inflammation by impacting the Th17/Treg balance in LPS-induced ARDS mice

“…Then, the cells were cultured in serum-free DMEM/F12 for another 12 h. The images of the wound area were recorded by a light microscope immediately after scratching and 12 h later. The horizontal migration ability of the cells was quantified by measuring the wound area in each group by Image J analysis software [13].…”

“…After incubation for 12 h, the cells remaining on the upper surface of the inserts were removed with cotton swabs, and the cells that had migrated to the lower surface were stained with crystal violet (Beyotime Institute of Biotechnology, Haimen, China) for 20 min. The stained cells from four randomly chosen areas were measured under a light microscope [13].…”

Overexpression of TGFβ1 in murine mesenchymal stem cells improves lung inflammation by impacting the Th17/Treg balance in LPS-induced ARDS mice

“…(Nanjing, China). The mBM-MSC were identi ed by detecting cell surface phenotypes and their multipotent potential for differentiation along the adipogenic, osteogenic, and chondrogenic lineages as previously described 13,14 .…”

Overexpression of TGFβ1 in murine mesenchymal stem cells improves lung inflammation by impacting the Th17/Treg balance in LPS-induced ARDS mice

“…Cell culture mMSCs isolated from the bone marrow of C57BL/6 mice (mBM-MSC) were purchased from Cyagen Biosciences, Inc. (Guangzhou, China), and 293T cells were supplied by Zoonbio Biotechnology Co., Ltd. (Nanjing, China). The mBM-MSC were identified by detecting cell surface phenotypes and their multipotent potential for differentiation along the adipogenic, osteogenic, and chondrogenic lineages as previously described 13,14 .…”

“…Then, the cells were cultured in serumfree DMEM/F12 for another 12 h. The images of the wound area were recorded by a light microscope immediately after scratching and 12 h later. The horizontal migration ability of the cells was quantified by measuring the wound area in each group by Image J analysis software 13 .…”