The positional identity of iPSC-derived neural progenitor cells along the anterior-posterior axis is controlled in a dosage-dependent manner by bFGF and EGF

Zhou, Shuling; Ochałek, Anna; Szczesna, Karolina; Avci, Hasan X.; Kobolák, Julianna; Varga, Eszter; Rasmussen, Mette; Holst, Bjørn; Cirera, Susanna; Hyttel, Poul; Freude, Kristine; Dinnyés, András

doi:10.1016/j.diff.2016.06.002

Cited by 8 publications

(5 citation statements)

References 47 publications

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“…FGF2 and EGF are growth factors that are known to stimulate differentiation of neural stem/progenitor cells (NSPCs). 25,26 Consistent with the reported effects, FGF2 and EGF treatment decreased the stemness of BMSCs as evidenced by the reduction of the percentages of CD54/CD90 positive cells and the reduced potential of BMSCs to be induced into adipocytes or osteocytes. In association with the differentiation, the elongated and interconnected mitochondria became dominant, as had been previously reported in P19 cells, embryonic stem cells, and hematopoietic stem cells.…”

Section: Discussionsupporting

confidence: 82%

“…FGF2 and EGF are growth factors that are known to stimulate differentiation of stem cells. 25,26 After treatment with FGF2 and EGF, cells with high levels of CD90 and CD54 (stemness markers) were decreased (Figure 2(a)). Correspondingly, the potency of BMSCs to be induced into osteocytes or adipocytes was also decreased significantly (Figure 2(b) and (c)), indicating a successful induction of differentiation state of BMSCs by FGF2 and EGF treatment.…”

Section: Resultsmentioning

confidence: 99%

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The involvement of mitochondrial fission in maintenance of the stemness of bone marrow mesenchymal stem cells

Feng

Zhang

Yin

et al. 2019

Exp Biol Med (Maywood)

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Mitochondrial dynamics, a complicated cellular process consisting of mitochondrial fusion and fission, has been suggested to be involved in regulating the stemness of bone marrow mesenchymal stem cells (BMSCs). This study was undertaken to explore the relationship between mitochondrial dynamics and the maintenance of BMSCs’ stemness. Rat BMSCs were treated with fibroblast growth factor 2 (FGF2) and epithelial growth factor (EGF) to induce differentiation. Mitochondrial dynamics was determined by mitochondrial length observed by confocal microscope and DLP1 (a protein promoting mitochondrial fission), OPA1 (a protein promoting mitochondrial fusion) expression revealed by Western blotting analysis. BMSCs’ stemness was determined by flow cytometry and osteogenic/adipogenic differentiation ability. We found that in the process of BMSCs differentiation, mitochondrial length was increased, along with a decreased protein level of DLP1 and an increased protein level of OPA1 in the mitochondria, indicating a shift toward mitochondrial fusion in BMSCs during differentiation. Notably, when the mitochondrial fission was inhibited by Mdivi-1, the stemness marker, CD90, was deceased along with the reduction of DLP1 expression. Under the same condition, the potential of BMSCs to be induced into adipocytes or osteocytes was decreased. Correspondingly, when BMSCs were treated with tyrphostin A9, a reagent promoting mitochondrial fission by increasing DLP1, the stemness marker, CD54, was increased with an increased potential of BMSCs to be induced into adipocytes or osteocytes. Hence, our results demonstrated that mitochondrial fission contributed to the maintenance of BMSCs’ stemness. Impact statement How to maintain the stemness of bone marrow mesenchymal stem cells (BMSCs) in cultures is a long-standing question. The present study found that mitochondrial dynamics affects the stemness of BMSCs in cultures and the retaining of mitochondrial fission enhances the stemness of BMSCs. This work thus provides a novel insight into strategic approaches to maintain the stemness of BMSCs in cultures in relation to the clinical application of bone-marrow stem cells.

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Section: Discussionsupporting

confidence: 82%

Section: Resultsmentioning

confidence: 99%

The involvement of mitochondrial fission in maintenance of the stemness of bone marrow mesenchymal stem cells

Feng

Zhang

Yin

et al. 2019

Exp Biol Med (Maywood)

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“…Our hiPSC-NPCs showed significant ventral hindbrain markers, which could have been induced ventrally by the inhibition of bone morphogenetic protein (BMP) signaling during the differentiation induction phase of the dual SMAD inhibition protocol and differentiation into the hindbrain system via the EGF and FGF used in the NPC culture. BMP signaling is known to promote dorsalization during the process of differentiation into neurons, and NPCs are thought to be transformed into hindbrain progenitor cells when EGF and FGF are added [39][40][41]. However, there have been reports of differentiation into the same ventral side of the brain that differentiates into GAD67-positive cells [9].…”

Section: In Vitro-expanded Hipsc-npcs Predominantly Differentiate Into Vglut2-positive Glutamatergic Neuronsmentioning

confidence: 99%

Postsynaptic structure formation of human iPS cell-derived neurons takes longer than presynaptic formation during neural differentiation in vitro

et al. 2021

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The generation of mature synaptic structures using neurons differentiated from human-induced pluripotent stem cells (hiPSC-neurons) is expected to be applied to physiological studies of synapses in human cells and to pathological studies of diseases that cause abnormal synaptic function. Although it has been reported that synapses themselves change from an immature to a mature state as neurons mature, there are few reports that clearly show when and how human stem cell-derived neurons change to mature synaptic structures. This study was designed to elucidate the synapse formation process of hiPSC-neurons. We propagated hiPSC-derived neural progenitor cells (hiPSC-NPCs) that expressed localized markers of the ventral hindbrain as neurospheres by dual SMAD inhibition and then differentiated them into hiPSC-neurons in vitro. After 49 days of in vitro differentiation, hiPSC-neurons significantly expressed pre- and postsynaptic markers at both the transcript and protein levels. However, the expression of postsynaptic markers was lower than in normal human or normal rat brain tissues, and immunostaining analysis showed that it was relatively modest and was lower than that of presynaptic markers and that its localization in synaptic structures was insufficient. Neurophysiological analysis using a microelectrode array also revealed that no synaptic activity was generated on hiPSC-neurons at 49 days of differentiation. Analysis of subtype markers by immunostaining revealed that most hiPSC-neurons expressed vesicular glutamate transporter 2 (VGLUT2). The presence or absence of NGF, which is required for the survival of cholinergic neurons, had no effect on their cell fractionation. These results suggest that during the synaptogenesis of hiPSC-neurons, the formation of presynaptic structures is not the only requirement for the formation of postsynaptic structures and that the mRNA expression of postsynaptic markers does not correlate with the formation of their mature structures. Technically, we also confirmed a certain level of robustness and reproducibility of our neuronal differentiation method in a multicenter setting, which will be helpful for future research. Synapse formation with mature postsynaptic structures will remain an interesting issue for stem cell-derived neurons, and the present method can be used to obtain early and stable quality neuronal cultures from hiPSC-NPCs.

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“…Among other cytokines which exhibit a significant increase, we found EGF, bFGF, AR (amphiregulin, a member of the EGF family), insulin, HGF (hepatocyte growth factor), OPG (Ooteoprotegrin), IGF (insulin-like growth factor), GDNF (glial cell linederived neurotrophic growth factor), etc. As we all know, EGF and bFGF have long been shown to play an important role in neural stem cell maintenance and proliferation [34,35]. The highly increased expression of EGF and bFGF means that these MSC-derived neurospheres can be selfsufficient.…”

mentioning

confidence: 99%

Efficient One-Step Induction of Human Umbilical Cord-Derived Mesenchymal Stem Cells (UC-MSCs) Produces MSC-Derived Neurospheres (MSC-NS) with Unique Transcriptional Profile and Enhanced Neurogenic and Angiogenic Secretomes

Peng

et al. 2019

Stem Cells International

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Cell therapy has emerged as a promising strategy for treating neurological diseases such as stroke, spinal cord injury, and various neurodegenerative diseases, but both embryonic neural stem cells and human induced Pluripotent Stem Cell- (iPSC-) derived neural stem cells have major limitations which restrict their broad use in these diseases. We want to find a one-step induction method to transdifferentiate the more easily accessible Umbilical Cord-Derived Mesenchymal Stem Cells (UC-MSCs) into neural stem/progenitor cells suitable for cell therapy purposes. In this study, UC-MSCs were induced to form neurospheres under a serum-free suspension culture with Epidermal Growth Factor- (EGF-) and basic Fibroblast Growth Factor- (bFGF-) containing medium within 12 hours. These MSC-derived neurospheres can self-renew to form secondary neurospheres and can be readily induced to become neurons and glial cells. Real-time PCR showed significantly upregulated expression of multiple stemness and neurogenic genes after induction. RNA transcriptional profiling study showed that UC-MSC-derived neurospheres had a unique transcriptional profile of their own, with features of both UC-MSCs and neural stem cells. RayBio human growth factor cytokine array analysis showed significantly upregulated expression levels of multiple neurogenic and angiogenic growth factors, skewing toward a neural stem cell phenotype. Thus, we believe that these UC-MSC-derived neurospheres have amenable features of both MSCs and neural stem/progenitor cells and have great potential in future stem cell transplantation clinical trials targeting neurological disorders.

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The positional identity of iPSC-derived neural progenitor cells along the anterior-posterior axis is controlled in a dosage-dependent manner by bFGF and EGF

Cited by 8 publications

References 47 publications

The involvement of mitochondrial fission in maintenance of the stemness of bone marrow mesenchymal stem cells

The involvement of mitochondrial fission in maintenance of the stemness of bone marrow mesenchymal stem cells

Postsynaptic structure formation of human iPS cell-derived neurons takes longer than presynaptic formation during neural differentiation in vitro

Efficient One-Step Induction of Human Umbilical Cord-Derived Mesenchymal Stem Cells (UC-MSCs) Produces MSC-Derived Neurospheres (MSC-NS) with Unique Transcriptional Profile and Enhanced Neurogenic and Angiogenic Secretomes

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