Biophysical Characteristics Reveal Neural Stem Cell Differentiation Potential

Labeed, Fatima H.; Lu, Juin-Ming; Mulhall, Hayley J.; Marchenko, Steve; Hoettges, Kai F.; Estrada, Laura C.; Lee, Abraham P.; Hughes, Michael; Flanagan, Lisa A.

doi:10.1371/journal.pone.0025458

Cited by 75 publications

(118 citation statements)

References 54 publications

Supporting

Mentioning

109

Contrasting

Order By: Relevance

“…The change in capacitance could also be related to changes in the permittivity of the membrane due to the action of 4-MU in shortening the HA chains; however, we suggest that of the two mechanisms, the effects of morphology charge are likely to significantly outweigh those of permittivity change. The unique differences in the membrane electrophysiological properties of stem-like cells is in line with a recent finding from our group showing that cell membrane dielectric properties could potentially be used as a marker for stem cells in neural tissues 39 . With a lack of distinct stem cell markers, isolation of tissuespecific stem cells for tissue engineering and gene therapy is a great challenge.…”

Section: Discussionsupporting

confidence: 87%

Human oral cancer cells with increasing tumorigenic abilities exhibit higher effective membrane capacitance

et al. 2014

View full text Add to dashboard Cite

Objective: Although cells with tumorigenic / stem cell-like properties have been identified in many cancers, including oral squamous cell carcinoma (OSCC), their isolation and characterisation is still at early stages. The aim of this study was to characterise the electrophysiological properties of OSCC cells with different tumorigenic properties in order to establish if a correlation exists between tumorigenicity and electrical characteristics. Materials and Methods: Rapid adherence to collagen IV was used as a non-invasive, functional method to isolate subsets of cells with different tumorigenic abilities from one oral dysplastic and three OSCC-derived cell lines. The cell subsets identified and isolated by this method were further investigated independently using dielectrophoresis, a label-free method to determine their electrophysiological parameters. Cell membrane morphology was investigated using scanning electron microscopy (SEM) and modulated by use of 4-methylumbelliferone (4-MU). Results: The rapid adherent cells (RAC) to collagen IV, enriched for cells with increased tumorigenic ability, had significantly higher effective membrane capacitance than middle (MAC) and late (LAC) adherent cells. SEM showed that, in contrast to MAC and LAC, RAC displayed a rough surface, extremely rich in cellular protrusions. Treatment with 4-MU dramatically altered RAC cell membrane morphology by causing loss of filopodia, and decreased significantly their membrane capacitance, indicating that the highest membrane capacitance found in RAC was due to their particular cell membrane morphology. Conclusion: This is the first study showing that OSCC cells with higher tumour formation ability exhibit higher effective membrane capacitance than cells that are less tumorigenic. OSSC cells with different tumorigenic ability possessed different electrophysiological properties mostly due to their differences in the cell membrane morphology. The results suggest that dielectrophoresis may potentially be of use in the future for reliable, label-free isolation of putative tumorigenic cells.

show abstract

Section: Discussionsupporting

confidence: 87%

Human oral cancer cells with increasing tumorigenic abilities exhibit higher effective membrane capacitance

et al. 2014

View full text Add to dashboard Cite

show abstract

“…Both cultures express standard neural stem cell markers, including Sox2, nestin, and the cell surface marker CD133, and have the potential to differentiate into the three major neural cell types: astrocytes, neurons, and oligodendrocytes (18)(19)(20)(21)(22). When transplanted into a mouse model of Sandhoff disease, these hNSPCs stably integrated into the host brain and delayed disease onset, reflecting their functional and therapeutic potential (23).…”

Section: Resultsmentioning

confidence: 99%

“…Patch clamp measurements, TIRFM Ca 2+ imaging, and differentiation assays were performed on two brain-derived human fetal hNSPC cultures (SC23 and SC27) isolated from the cerebral cortices of two separate fetuses of 23-wk gestational age (18)(19)(20)(21). Further details on cell-culture conditions, siRNA knockdown experiments, electrophysiological measurements, calcium imaging, Qgel substrate fabrication, and data analysis are presented in SI Methods.…”

Section: Methodsmentioning

confidence: 99%

Stretch-activated ion channel Piezo1 directs lineage choice in human neural stem cells

Pathak¹,

Nourse

Tran³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

502

509

View full text Add to dashboard Cite

Neural stem cells are multipotent cells with the ability to differentiate into neurons, astrocytes, and oligodendrocytes. Lineage specification is strongly sensitive to the mechanical properties of the cellular environment. However, molecular pathways transducing matrix mechanical cues to intracellular signaling pathways linked to lineage specification remain unclear. We found that the mechanically gated ion channel Piezo1 is expressed by brainderived human neural stem/progenitor cells and is responsible for a mechanically induced ionic current. Piezo1 activity triggered by traction forces elicited influx of Ca 2+ , a known modulator of differentiation, in a substrate-stiffness-dependent manner. Inhibition of channel activity by the pharmacological inhibitor GsMTx-4 or by siRNA-mediated Piezo1 knockdown suppressed neurogenesis and enhanced astrogenesis. Piezo1 knockdown also reduced the nuclear localization of the mechanoreactive transcriptional coactivator Yes-associated protein. We propose that the mechanically gated ion channel Piezo1 is an important determinant of mechanosensitive lineage choice in neural stem cells and may play similar roles in other multipotent stem cells. (5) and that the mechanical properties of the culturing environment before transplantation can influence the outcome of in vivo stem cell transplants (6). Hence, a molecular and mechanistic understanding of how stem cells process mechanical cues and how this processing results in downstream signaling events and ultimately in fate decisions is needed for greater control over the fate of transplanted cells.Studies in mesenchymal and neural stem cells have revealed the involvement of focal adhesion zones and cytoskeletal proteins, such as integrins, nonmuscle myosin II (7), Rho GTPases (8-10), and vinculin (11), that participate in the generation of cellular traction forces. Recent work also has identified the nucleoskeletal protein lamin-A (12) and the transcriptional coactivators Yap (Yesassociated protein) and Taz (transcriptional coactivator with PDZbinding motif) (13) in mechanotransduction in mesenchymal stem cells. However, the mechanisms by which mechanical cues detected by cellular traction forces are transduced to downstream intracellular pathways of differentiation remain unclear.Ion channels are involved, directly or indirectly, in the transduction of all forms of physical stimuli-including sound, light, temperature, mechanical force, and even gravity-into intracellular signaling pathways. Hence, we wondered whether ion channels could be involved in transducing matrix mechanical cues to intracellular signaling pathways linked to lineage specification. In particular, we focused here on cationic stretch-activated channels (SACs) because they are known to detect mechanical forces with high sensitivity and broad dynamic range and because they are permeable to Ca 2+ , an important second messenger implicated in cell fate (14,15). We examined the role of SACs in neural stem cells, for which mechanical cues influence specification alo...

show abstract

“…(2) leads to an accurate measurement of the membrane capacitance. 16 Labeed et al 17 have recently reported that the neurogenic potential of human neural stem/progenitor cell populations can be discriminated by their f xo and derived C m values. On the basis of the efficient DEP-based cell separations reported here, the same conclusion may be reached regarding discrimination between stages of C2C12 myoblast cell differentiation.…”

Section: -1058/2012/6(3)/034113/16/$3000mentioning

confidence: 99%

Biomarker-free dielectrophoretic sorting of differentiating myoblast multipotent progenitor cells and their membrane analysis by Raman spectroscopy

et al. 2012

View full text Add to dashboard Cite

Myoblasts are muscle derived mesenchymal stem cell progenitors that have great potential for use in regenerative medicine, especially for cardiomyogenesis grafts and intracardiac cell transplantation. To utilise such cells for pre-clinical and clinical applications, and especially for personalized medicine, it is essential to generate a synchronised, homogenous, population of cells that display phenotypic and genotypic homogeneity within a population of cells. We demonstrate that the biomarker-free technique of dielectrophoresis (DEP) can be used to discriminate cells between stages of differentiation in the C2C12 myoblast multipotent mouse model. Terminally differentiated myotubes were separated from C2C12 myoblasts to better than 96% purity, a result validated by flow cytometry and Western blotting. To determine the extent to which cell membrane capacitance, rather than cell size, determined the DEP response of a cell, C2C12 myoblasts were co-cultured with GFP-expressing MRC-5 fibroblasts of comparable size distributions (mean diameter $10 lm). A DEP sorting efficiency greater than 98% was achieved for these two cell types, a result concluded to arise from the fibroblasts possessing a larger membrane capacitance than the myoblasts. It is currently assumed that differences in membrane capacitance primarily reflect differences in the extent of folding or surface features of the membrane. However, our finding by Raman spectroscopy that the fibroblast membranes contained a smaller proportion of saturated lipids than those of the myoblasts suggests that the membrane chemistry should also be taken into account.

show abstract

Biophysical Characteristics Reveal Neural Stem Cell Differentiation Potential

Cited by 75 publications

References 54 publications

Human oral cancer cells with increasing tumorigenic abilities exhibit higher effective membrane capacitance

Human oral cancer cells with increasing tumorigenic abilities exhibit higher effective membrane capacitance

Stretch-activated ion channel Piezo1 directs lineage choice in human neural stem cells

Biomarker-free dielectrophoretic sorting of differentiating myoblast multipotent progenitor cells and their membrane analysis by Raman spectroscopy

Contact Info

Product

Resources

About