scholarly journals Nanobiosensing Platforms for Real-time and Non-Invasive Monitoring of Stem Cell Pluripotency and Differentiation

Sensors ◽  
2018 ◽  
Vol 18 (9) ◽  
pp. 2755 ◽  
Author(s):  
Intan Rosalina Suhito ◽  
Novi Angeline ◽  
Sung-Sik Choo ◽  
Ho Young Woo ◽  
Taejong Paik ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Stem Cell Differentiation ◽  
Electrochemical Sensing ◽  
Non Invasive ◽  
Differentiated Cells ◽  
Present Information ◽  
Stem Cell Pluripotency ◽  
Technical Motivation ◽  
Non Destructive

Breakthroughs in the biomedical and regenerative therapy fields have led to the influential ability of stem cells to differentiate into specific types of cells that enable the replacement of injured tissues/organs in the human body. Non-destructive identification of stem cell differentiation is highly necessary to avoid losses of differentiated cells, because most of the techniques generally used as confirmation tools for the successful differentiation of stem cells can result in valuable cells becoming irrecoverable. Regarding this issue, recent studies reported that both Raman spectroscopy and electrochemical sensing possess excellent characteristics for monitoring the behavior of stem cells, including differentiation. In this review, we focus on numerous studies that have investigated the detection of stem cell pluripotency and differentiation in non-invasive and non-destructive manner, mainly by using the Raman and electrochemical methods. Through this review, we present information that could provide scientific or technical motivation to employ or further develop these two techniques for stem cell research and its application.

scholarly journals Epigenetics, Stem Cells, and Autophagy: Exploring a Path Involving miRNA

2019 ◽  
Vol 20 (20) ◽  
pp. 5091 ◽  
Author(s):  
Francesca Balzano ◽  
Ilaria Campesi ◽  
Sara Cruciani ◽  
Giuseppe Garroni ◽  
Emanuela Bellu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Dna Methyltransferase ◽  
Main Tool ◽  
Stem Cell Differentiation ◽  
Differentiation Potential ◽  
Multipotent Stem Cells ◽  
Regulatory Circuit ◽  
Stem Cell Pluripotency ◽  
Males And Females

MiRNAs, a small family of non-coding RNA, are now emerging as regulators of stem cell pluripotency, differentiation, and autophagy, thus controlling stem cell behavior. Stem cells are undifferentiated elements capable to acquire specific phenotype under different kind of stimuli, being a main tool for regenerative medicine. Within this context, we have previously shown that stem cells isolated from Wharton jelly multipotent stem cells (WJ-MSCs) exhibit gender differences in the expression of the stemness related gene OCT4 and the epigenetic modulator gene DNA-Methyltransferase (DNMT1). Here, we further analyze this gender difference, evaluating adipogenic and osteogenic differentiation potential, autophagic process, and expression of miR-145, miR-148a, and miR-185 in WJ-MSCs derived from males and females. These miRNAs were selected since they are involved in OCT4 and DNMT1 gene expression, and in stem cell differentiation. Our results indicate a difference in the regulatory circuit involving miR-148a/DNMT1/OCT4 autophagy in male WJ-MSCs as compared to female cells. Moreover, no difference was detected in the expression of the two-differentiation regulating miRNA (miR-145 and miR-185). Taken together, our results highlight a different behavior of WJ-MSCs from males and females, disclosing the chance to better understand cellular processes as autophagy and stemness, usable for future clinical applications.

Identification of mesenchymal stem cell differentiation state using dual-micropore microfluidic impedance flow cytometry

2016 ◽  
Vol 8 (41) ◽  
pp. 7437-7444 ◽  
Author(s):  
Hongjun Song ◽  
Jenna M. Rosano ◽  
Yi Wang ◽  
Charles J. Garson ◽  
Balabhaskar Prabhakarpandian ◽  
...  
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Cell Differentiation ◽  
Mesenchymal Stem Cell ◽  
Electrical Impedance ◽  
Stem Cell Differentiation ◽  
Non Invasive ◽  
Mesenchymal Stem Cell Differentiation

A dual-micropore-based microfluidic electrical impedance flow cytometer for non-invasive identification of the differentiation state of mesenchymal stem cells.

scholarly journals A secreted proteomic footprint for stem cell pluripotency

2021 ◽  
Author(s):  
Philip A Lewis ◽  
Edina Silajdžić ◽  
Helen Smith ◽  
Nicola Bates ◽  
Christopher A Smith ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Pluripotent Stem Cells ◽  
Strong Correlation ◽  
Proteomic Analysis ◽  
Human Stem Cells ◽  
Pluripotent State ◽  
Non Invasive ◽  
Stem Cell Pluripotency ◽  
Invasive Method

With a view to developing a much-needed non-invasive method for monitoring the healthy pluripotent state of human stem cells in culture, we undertook proteomic analysis of the spent medium from cultured embryonic (Man-13) and induced (Rebl.PAT) human pluripotent stem cells (hPSCs). Cells were grown in E8 medium to maintain pluripotency, and then transferred to FGF2 and TGFβ deficient media for 48 hours to replicate an early, undirected dissolution of pluripotency. We identified a distinct proteomic footprint associated with early loss of pluripotency in both hPSC lines, and a strong correlation with changes in the transcriptome. We demonstrate that multiplexing of 4 E8- against 4 E6- enriched biomarkers provides 16 ratio abundances which are each robustly diagnostic for pluripotent state. These biomarkers were further confirmed by Western blotting which demonstrated consistent correlation with the pluripotent state across cell lines, and in response to recovery assays.

scholarly journals Recognition of tenogenic differentiation using convolutional neural network

2020 ◽  
Vol 6 (3) ◽  
pp. 200-204
Author(s):  
Gözde Dursun ◽  
Saurabh Balkrishna Tandale ◽  
Jörg Eschweiler ◽  
Mersedeh Tohidnezhad ◽  
Bernd Markert ◽  
...  
Keyword(s):  
Neural Networks ◽  
Stem Cells ◽  
Stem Cell ◽  
Cell Differentiation ◽  
High Performance ◽  
Image Data ◽  
Stem Cell Differentiation ◽  
Differentiation Potential ◽  
Non Invasive ◽  
Invasive Cell

AbstractMethodologies to assess stem cell differentiation in the culturing state are needed for regenerative medicine and tissue engineering techniques. In recent years, convolutional neural networks (CNNs), a class of deep neural networks, have made impressive advancements in image-based classification, recognition and detection tasks. CNNs have been introduced as a non-invasive cell characterization method by learning features directly from image data of unlabeled cells. Furthermore, this approach serves as a rapid and inexpensive methodology with high performance compared to traditional techniques that require complex laboratory procedures including antibody staining and gene expression analysis. Here, we studied the potential of the CNNs approach to recognize stem cell differentiation based on cell morphology utilizing phasecontrast microscopy images.We have examined the differentiation potential of bone marrow mesenchymal stem cells (BMSCs) into tenocytes, with the treatment of bone morphogenetic protein-12 (BMP-12). After treatment, the phase-contrast images of cells were obtained directly from cell culture flasks to train CNN and the differentiated phenotype of stem cells was characterized by immunostaining. CNN was able to classify the cells into three groups including non-stem cells (chondrocytes), stem cells (BMSCs) and differentiated stem cells (tenocytes) based on their morphology with 92.2 % accuracy. The presented study revealed that CNN performed faster and non-invasive cell classification task compared to traditional methodologies.

scholarly journals G-quadruplex DNA structures in human stem cells and differentiation

2022 ◽  
Vol 13 (1) ◽  
Author(s):  
Katherine G. Zyner ◽  
Angela Simeone ◽  
Sean M. Flynn ◽  
Colm Doyle ◽  
Giovanni Marsico ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Structural Features ◽  
Specific Gene ◽  
Human Stem Cells ◽  
Post Translational Modification ◽  
Cellular Functions ◽  
Stem Cell Pluripotency

AbstractThe establishment of cell identity during embryonic development involves the activation of specific gene expression programmes and is underpinned by epigenetic factors including DNA methylation and histone post-translational modifications. G-quadruplexes are four-stranded DNA secondary structures (G4s) that have been implicated in transcriptional regulation and cancer. Here, we show that G4s are key genomic structural features linked to cellular differentiation. We find that G4s are highly abundant in human embryonic stem cells and are lost during lineage specification. G4s are prevalent in enhancers and promoters. G4s that are found in common between embryonic and downstream lineages are tightly linked to transcriptional stabilisation of genes involved in essential cellular functions as well as transitions in the histone post-translational modification landscape. Furthermore, the application of small molecules that stabilise G4s causes a delay in stem cell differentiation, keeping cells in a more pluripotent-like state. Collectively, our data highlight G4s as important epigenetic features that are coupled to stem cell pluripotency and differentiation.

scholarly journals Non-destructive characterisation of mesenchymal stem cell differentiation using LC-MS-based metabolite footprinting

The Analyst ◽  
2016 ◽  
Vol 141 (12) ◽  
pp. 3776-3787 ◽  
Author(s):  
Amal Surrati ◽  
Rob Linforth ◽  
Ian D. Fisk ◽  
Virginie Sottile ◽  
Dong-Hyun Kim
Keyword(s):  
Stem Cell ◽  
Cell Differentiation ◽  
Mesenchymal Stem Cell ◽  
Stem Cell Differentiation ◽  
Metabolic Changes ◽  
Non Invasive ◽  
Mesenchymal Stem Cell Differentiation ◽  
Non Destructive ◽  
Msc Differentiation

This non-invasive LC-MS-based metabolite footprinting revealed significant metabolic changes during mesenchymal stem cell (MSC) differentiation.

Oct4 and Its Pseudogenes Confuse Stem Cell Research.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3700-3700
Author(s):  
Stefanie Liedtke ◽  
Jürgen Enczmann ◽  
Simon Waclawczyk ◽  
Peter Wernet ◽  
Gesine Kögler
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
Adult Stem Cells ◽  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Differentiated Cells ◽  
Alternatively Spliced

Abstract Octamer-binding transcription factor 4 (Oct4) encodes a nuclear protein that belongs to a family of transcription factors containing the POU DNA binding domain. It is specifically expressed in embryonic stem cells but can also be detected in adult stem cells such as bone marrow-derived mesenchymal stem cells. The expression of Oct4 is down-regulated coincident with stem cell differentiation and loss of expression leading to differentiation. It plays a critical role for maintaining pluripotency and self-renewal of embryonic stem cells. However, the usefulness of Oct4 as a pluripotency marker was challenged recently. More and more data seem to support that Oct4 is expressed on a variety of differentiated cells, including peripheral blood mononuclear cells. Taking into account that RT-PCR can potentially generate experimental artifacts due to pseudogene transcripts, the existence of Oct4 pseudogenes should be investigated further here. Suo et al. were able to detect transcription of some Oct4 pseudogenes in cancer cell lines as well as cancer tissues. These results show that some of the known Oct4 pseudogenes are transcribed in vivo and therefore could lead to RT-PCR artifacts. However this known problem was not seriously taken into consideration in recent publications on adult stem cells and tissue analysis referring to Oct4. We started with an initial alignment of Oct4 compared to its alternative splice variants as well as its pseudogenes. This alignment served as a prerequisite for an exact primer design. First the sequence and organization of the functional human Oct4 gene were clarified to allow comparison to the pseudogenes and alternatively spliced transcripts. The NCBI human EST database was searched and the UniGene cluster for Oct4 (NM_002701) examined. This yielded 13 mRNA sequences and 129 EST sequences. An additional BLASTn search of the human genome using single exons of Oct4 revealed several other highly similar sequences. All these hits encoded complete or partial Oct4 sequences and could therefore represent either functional members of an Oct4 gene family or pseudogenes. The fact that so many homologous sequences resemble the original Oct4 transcript makes an RT-PCR analysis difficult, because a lot of artifacts can arise during amplification. Therefore primers were designed which are able to exclude amplification of all unwanted transcripts. To conclude, based on the fact that the expression of Oct4 has been reported in adult stem cells as well as in a variety of differentiated cells the possibility cannot be excluded that the detected Oct4 signal came from alternatively spliced or Oct4 pseudogene transcripts. As shown here, an exact design of Oct4-specific primers is an inevitable prerequisite for appropriate RT-PCR analysis. In addition, a careful comparison of quantitative differences to human embryonic stem cells should be present too, before cells are described as embryonic like cells. We hope that our findings will help other stem cell researchers to find their appropriate tools especially for RT-PCR analysis and give an example how later problematic artifacts can be ruled out from the beginning by a detailed alignment as a prerequisite for designing appropriate primers.

scholarly journals DYRK1A Negatively Regulates CDK5-SOX2 Pathway and Self-Renewal of Glioblastoma Stem Cells

2021 ◽  
Vol 22 (8) ◽  
pp. 4011
Author(s):  
Brianna Chen ◽  
Dylan McCuaig-Walton ◽  
Sean Tan ◽  
Andrew P. Montgomery ◽  
Bryan W. Day ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Cell Differentiation ◽  
Critical Role ◽  
Stem Cell Differentiation ◽  
Neural Progenitor Cell ◽  
Cellular Heterogeneity ◽  
Differentiation Potential ◽  
Glioblastoma Stem Cells ◽  
Glioblastoma Stem Cell

Glioblastoma display vast cellular heterogeneity, with glioblastoma stem cells (GSCs) at the apex. The critical role of GSCs in tumour growth and resistance to therapy highlights the need to delineate mechanisms that control stemness and differentiation potential of GSC. Dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A) regulates neural progenitor cell differentiation, but its role in cancer stem cell differentiation is largely unknown. Herein, we demonstrate that DYRK1A kinase is crucial for the differentiation commitment of glioblastoma stem cells. DYRK1A inhibition insulates the self-renewing population of GSCs from potent differentiation-inducing signals. Mechanistically, we show that DYRK1A promotes differentiation and limits stemness acquisition via deactivation of CDK5, an unconventional kinase recently described as an oncogene. DYRK1A-dependent inactivation of CDK5 results in decreased expression of the stemness gene SOX2 and promotes the commitment of GSC to differentiate. Our investigations of the novel DYRK1A-CDK5-SOX2 pathway provide further insights into the mechanisms underlying glioblastoma stem cell maintenance.

scholarly journals Nuclear Export of Smads by RanBP3L Regulates Bone Morphogenetic Protein Signaling and Mesenchymal Stem Cell Differentiation

2015 ◽  
Vol 35 (10) ◽  
pp. 1700-1711 ◽  
Author(s):  
Fenfang Chen ◽  
Xia Lin ◽  
Pinglong Xu ◽  
Zhengmao Zhang ◽  
Yanzhen Chen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Cell Differentiation ◽  
Mesenchymal Stem Cell ◽  
Nuclear Export ◽  
Stem Cell Differentiation ◽  
Bmp Signaling ◽  
Dependent Manner ◽  
Mesenchymal Stem Cell Differentiation

Bone morphogenetic proteins (BMPs) play vital roles in regulating stem cell maintenance and differentiation. BMPs can induce osteogenesis and inhibit myogenesis of mesenchymal stem cells. Canonical BMP signaling is stringently controlled through reversible phosphorylation and nucleocytoplasmic shuttling of Smad1, Smad5, and Smad8 (Smad1/5/8). However, how the nuclear export of Smad1/5/8 is regulated remains unclear. Here we report that the Ran-binding protein RanBP3L acts as a nuclear export factor for Smad1/5/8. RanBP3L directly recognizes dephosphorylated Smad1/5/8 and mediates their nuclear export in a Ran-dependent manner. Increased expression of RanBP3L blocks BMP-induced osteogenesis of mouse bone marrow-derived mesenchymal stem cells and promotes myogenic induction of C2C12 mouse myoblasts, whereas depletion of RanBP3L expression enhances BMP-dependent stem cell differentiation activity and transcriptional responses. In conclusion, our results demonstrate that RanBP3L, as a nuclear exporter for BMP-specific Smads, plays a critical role in terminating BMP signaling and regulating mesenchymal stem cell differentiation.

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