Isolation and characterization of neural progenitor cells from post‐mortem human cortex

Schwartz, Philip H.; Bryant, Peter J.; Fuja, Tannin J.; Su, Hailing; O’Dowd, Diane K.; Klassen, Henry

doi:10.1002/jnr.10854

Cited by 237 publications

(255 citation statements)

References 46 publications

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“…We used cultures of the hNSPCs SC23 and SC27 derived from the cerebral cortices of two separate postmortem fetal brains (18,19) to test the role of SACs in mechanical modulation of stem cell behavior. 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).…”

Section: Resultsmentioning

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.

483

509

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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...

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Section: Resultsmentioning

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.

483

509

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“…The human gliomas U87, U251, D54, LN18, LN229, T98G, and LNZ308 were described in Schwartz et al (26). The HLA typing provides evidence of the genetic differences of these cells.…”

Section: Cell Lines and Surgical Specimensmentioning

confidence: 99%

“…The stem cells were cultured cells on Matrigel-coated dishes with DMEM/F12 (1:1 v/v) medium (Irvine Scientific, Irvine, CA) containing 10% BIT 9500 (BSA, insulin, transferrin; Stem Cell Technologies, Tukwila, WA), 292 mg/ml glutamine (Irvine Scientific), 40 ng/ml basic fibroblast growth factor, 20 ng/ml epidermal growth factor, and 20 ng/ml platelet-derived growth factor-AB, 100 U/ml penicillin (Irvine Scientific), 100 mg/ml streptomycin (Irvine Scientific), 50 mg/ml gentamicin (Sigma-Aldrich), 10 mg/ml ciprofloxacin (Bayer), and 2.5 mg/ml amphotericin (Life Technologies/Invitrogen, Carlsbad, CA) as previously described (26). For continuous expansion, half of this medium was replaced every other day, and the cultures were passaged every 7 d or when confluent using nonenzymatic cell dissociation solution (Sigma-Aldrich).…”

Section: Normal Human Brain Neurosphere Culturesmentioning

confidence: 99%

Glioma Big Potassium Channel Expression in Human Cancers and Possible T Cell Epitopes for Their Immunotherapy

Ge¹,

Hoa²,

Cornforth

et al. 2012

The Journal of Immunology

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Big potassium (BK) ion channels have several spliced variants. One spliced variant initially described within human glioma cells is the glioma BK (gBK) channel. This isoform consists of 34 aa inserted into the intracellular region of the basic BK ion channel. PCR primers specific for this inserted region confirmed that human glioma cell lines and freshly resected surgical tissues from glioblastoma multiforme patients strongly expressed gBK mRNA. Normal human brain tissue very weakly expressed this transcript. An Ab specific for this gBK isoform confirmed that human glioma cells displayed this protein in the cell membrane, mitochondria, Golgi, and endoplasmic reticulum. Within the gBK region, two putative epitopes (gBK1 and gBK2) are predicted to bind to the HLA-A*0201 molecule. HLA-A*0201–restricted human CTLs were generated in vitro using gBK peptide-pulsed dendritic cells. Both gBK1 and gBK2 peptide-specific CTLs killed HLA-A2+/gBK+ gliomas, but they failed to kill non-HLA-A2–expressing but gBK+ target cells in cytolytic assays. T2 cells loaded with exogenous gBK peptides, but not with the influenza M1 control peptide, were only killed by their respective CTLs. The gBK-specific CTLs also killed a variety of other HLA-A*0201+ cancer cells that possess gBK, as well as HLA-A2+ HEK cells transfected with the gBK gene. Of clinical relevance, we found that T cells derived from glioblastoma multiforme patients that were sensitized to the gBK peptide could also kill target cells expressing gBK. This study shows that peptides derived from cancer-associated ion channels maybe useful targets for T cell-mediated immunotherapy.

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“…bNSC gene expression versus that of eNSCs. The former were derived from human brain [103] and the latter were derived from ePSCs [23]. Both cell types were cultured in bFGF/EGFcontaining media.…”

Section: Tumorigenicity Of Final Product-one Of the Basic Properties mentioning

confidence: 99%

Differentiation of neural lineage cells from human pluripotent stem cells

Schwartz

Brick

Stover

et al. 2008

Methods

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Human pluripotent stem cells have the unique properties of being able to proliferate indefinitely in their undifferentiated state and to differentiate into any somatic cell type. These cells are thus posited to be extremely useful for furthering our understanding of both normal and abnormal human development, providing a human cell preparation that can be used to screen for new reagents or therapeutic agents, and generating large numbers of differentiated cells that can be used for transplantation purposes. Critical among the applications for the latter are diseases and injuries of the nervous system, medical approaches to which have been, to date, primarily palliative in nature. Differentiation of human pluripotent stem cells into cells of the neural lineage, therefore, has become a central focus of a number of laboratories. This has resulted in the description in the literature of several dozen methods for neural cell differentiation from human pluripotent stem cells. Among these are methods for the generation of such divergent neural cells as dopaminergic neurons, retinal neurons, ventral motoneurons, and oligodendroglial progenitors. In this review, we attempt to fully describe most of these methods, breaking them down into five basic subdivisions: 1) starting material, 2) induction of loss of pluripotency, 3) neural induction, 4) neural maintenance and expansion, and 5) neuronal/glial differentiation. We also show data supporting the concept that undifferentiated human pluripotent stem cells appear to have an innate neural differentiation potential. In addition, we evaluate data comparing and contrasting neural stem cells differentiated from human pluripotent stem cells with those derived directly from the human brain.

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Isolation and characterization of neural progenitor cells from post‐mortem human cortex

Cited by 237 publications

References 46 publications

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

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

Glioma Big Potassium Channel Expression in Human Cancers and Possible T Cell Epitopes for Their Immunotherapy

Differentiation of neural lineage cells from human pluripotent stem cells

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