Isolation of rat Schwann cells based on cell sorting

Shen, Mi; Tang, Wei; Cao, Zheng; Cao, Xianglin; Ding, Fei

doi:10.3892/mmr.2017.6777

Cited by 8 publications

(8 citation statements)

References 25 publications

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“…Cultures were performed using three media solutions: DMEM, NB medium, and EBS, all with 10% FBS and 1% penicillin‐streptomycin. ShC cultures were performed using two media solutions of DMEM and EBS, all with 10% FBS and 1% penicillin‐streptomycin, as per convention (Pena et al., 2019; Shen, Tang, Cao, Cao, & Ding, 2017). Explant cultures were performed in solutions of NB Media and EBS solutions supplemented with B27 and 0.5 mM Glutamax as per convention so as to provide long‐term viability to maturing neurons (Brewer, Torricelli, Evege, & Price, 1993; Chen et al., 2010; Thomson et al., 2008).…”

Section: Methodsmentioning

confidence: 99%

Cerebrospinal fluid replacement solutions promote neuroglia migratory behaviors and spinal explant outgrowth in microfluidic culture

Cliver

Ayers

Brady

et al. 2020

J Tissue Eng Regen Med

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Disorders of the nervous system (NS) impact millions of adults, worldwide, as a consequence of traumatic injury, genetic illness, or chronic health conditions. Contemporary studies have begun to incorporate neuroglia into emerging NS therapies to harness the regenerative potential of glial‐mediated synapses in the brain and spinal cord. However, the role of cerebrospinal fluid (CSF) that surrounds neuroglia and interfaces with their associated synapses remains only partially explored. The flow of CSF within subarachnoid spaces (SAS) circulates essential polypeptides, metabolites, and growth factors that directly impact neural response and recovery via signaling with healthy glia. Despite the availability of artificial CSF solutions used in neurosurgery and NS treatments, tissue engineering projects continue to use cell culture media, such as Neurobasal (NB) and Dulbecco's Modified Eagle Medium (DMEM), for development and characterization of many transplantable cells, matrixes, and integrated cellular systems. The current study examined in vitro behaviors of glial Schwann cells (ShC) and spinal cord explants (SCE) within a CSF replacement solution, Elliott's B Solution (EBS), used widely in the treatment of NS disorders. Our tests used EBS to create defined chemical microenvironments of extracellular factors within a glial line (gLL) microfluidic device, previously described by our group. The gLL is comparable in scale to the in vivo SAS that envelopes endogenous CSF and enables molecular transport via mechanisms of convective diffusion. Our results illustrate that EBS solutions facilitate ShC survival, morphology, and proliferation similar to those measured in traditional DMEM, and additionally support glial chemotactic behaviors in response to brain‐derived growth factor (BDNF). Our data indicates that ShC undergo significant chemotaxis toward high and low concentration gradients of BDNF with statistical differences between gradients formed within diluents of EBS and DMEM solutions. Moreover, SCE cultured with EBS solutions facilitated measurement of neurite explant extension commensurate with reported in vivo measurements. This data highlights the translational significance and advantages of incorporating CSF replacement fluids to interrogate cellular behaviors and advance regenerative NS therapies.

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Section: Methodsmentioning

confidence: 99%

Cerebrospinal fluid replacement solutions promote neuroglia migratory behaviors and spinal explant outgrowth in microfluidic culture

Cliver

Ayers

Brady

et al. 2020

J Tissue Eng Regen Med

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“…Consistent with the phenotypic changes, SV40T increased the expression levels of the neural crest markers Slug and Pax3, as well as the NSC marker Nestin in the SCs. In contrast, the SC marker S100b (Shen et al, 2017), and the markers of mature SCs MBP, GFAP and Olig1/2 were weakly expressed in these cells (Figure 3). Furthermore, the SV40T-SCs were positive for the NSC markers Nestin, Sox2, CD133 and SSEA-1, as well as the embryonic stem cell (ESC) markers BMP4, c-Myc, OCT4 and Gbx2 (Figure 4).…”

Section: Sv40t Reprogrammed the Scs Into Nsc-like Cellsmentioning

confidence: 95%

SV40T reprograms Schwann cells into stem-like cells that can re-differentiate into terminal nerve cells

Li¹,

Nan²,

Wang³

et al. 2021

Int. J. Dev. Biol.

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Background: The specific effect of SV40T on neurocytes has been rarely investigated by the researchers. We transfected Schwann cells (SCs) that did not have differentiation ability with MPH 86 plasmid containing SV40T in order to explore the effects of SV40T on Schwann cells.Methods: SCs were transfected with MPH 86 plasmid carrying the SV40T gene and cultured in different media, as well as co-cultured with neural stem cells (NSCs). In our study, SCs overexpressing SV40T were defined as SV40T-SCs. The proliferation of these cells was detected by WST-1, and the expression of different biomarkers was analyzed by qPCR and immunohistochemistry. Results: SV40T induced the characteristics of NSCs, such as the ability to grow in suspension, form spheroid colonies and proliferate rapidly, in the SCs, which were reversed by knocking out SV40T by the Flip-adenovirus. In addition, SV40T upregulated the expressions of neural crest-associated markers Nestin, Pax3 and Slug, and down-regulated S100b as well as the markers of mature SCs MBP, GFAP and Olig1/2. These cells also expressed NSC markers like Nestin, Sox2, CD133 and SSEA-1, as well as early development markers of embryonic stem cells (ESCs) like BMP4, c-Myc, OCT4 and Gbx2. Co-culturing with NSCs induced differentiation of the SV40T-SCs into neuronal and glial cells. Conclusions: SV40T reprograms Schwann cells to stem-like cells at the stage of neural crest cells (NCCs) that can differentiate to neurocytes.

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“…Consistent with the phenotypic changes, SV40T increased the expression levels of the neural crest markers Slug and Pax3, as well as the NSC marker Nestin in the SCs. In contrast, the SC marker S100b [12], and the later neuronal differentiation markers MBP, GFAP and Olig1/2 were weakly expressed in these cells ( Figure 3). Furthermore, the SV40T-SCs were positive for the NSC markers Nestin, Sox2, CD133 and SSEA-1, as well as the embryonic stem cell (ESC) markers BMP4, c-Myc, OCT4 and Gbx2 (Figure 4).…”

Section: Sv40t Reprogrammed the Scs Into Nsc-like Cellsmentioning

confidence: 97%

SV40T reprograms Schwann cells into stem-like cells that can re-differentiate into terminal nerve cells.

Nan

Wang

et al. 2020

Preprint

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Background : The effects of Simian virus 40 T antigen (SV40T) on various kinds of cells are different. Previous researchers failed to use SV40T immortalized nerve cells. However, they argued that SV40T caused nerve cell transformation. No one further study what is the specific effect of SV40T on nerve cells. We transfected Schwann cells (SCs) that did not have differentiation ability with MPH 86 plasmid containing SV40T in order to explore the effects of SV40T on Schwann cells. Methods : SCs were transfected with MPH 86 plasmid carrying the SV40T gene and cultured in different media, as well as co-cultured with neural stem cells (NSCs). In our study, SCs overexpressing SV40T were defined as SV40T-SCs. The proliferation of these cells was detected by WST-1, and the expression of different biomarkers was analyzed by qPCR and immunohistochemistry. Results: SV40T induced the characteristics of NSCs, such as the ability to grow in suspension, form spheroid colonies and proliferate rapidly, in the SCs, which were reversed by knocking out SV40T by the Flip-adenovirus . In addition, SV40T up-regulated the neurocrest markers Nestin, Pax3 and Slug, and down-regulated S100b as well as the late differentiation markers MBP, GFAP and Olig1/2. These cells also expressed NSC markers like Nestin, Sox2, CD133 and SSEA-1, as well as early development markers of embryonic stem cells (ESCs) like BMP4, c-Myc, OCT4 and Gbx2. Co-culturing with NSCs induced differentiation of the SV40T-SCs into neuronal and glial cells. Conclusions: SV40T reprograms Schwann cells to stem-like cells at the stage of neural crest cells that can differentiate to terminal nerve cells.

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Isolation of rat Schwann cells based on cell sorting

Cited by 8 publications

References 25 publications

Cerebrospinal fluid replacement solutions promote neuroglia migratory behaviors and spinal explant outgrowth in microfluidic culture

Cerebrospinal fluid replacement solutions promote neuroglia migratory behaviors and spinal explant outgrowth in microfluidic culture

SV40T reprograms Schwann cells into stem-like cells that can re-differentiate into terminal nerve cells

SV40T reprograms Schwann cells into stem-like cells that can re-differentiate into terminal nerve cells.

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