Novel method to obtain highly enriched cultures of adult rat Schwann cells

Niapour, Ali; Karamali, Fereshteh; Karbalaie, Khadijeh; Kiani, Abbas; Mardani, Mohammad; Nasr-Esfahani, Mohammad Hossein; Baharvand, Hossein

doi:10.1007/s10529-010-0230-z

Cited by 28 publications

(33 citation statements)

References 23 publications

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“…However, it is important to note that this is not a comparative study. Peripheral nerve predegeneration takes advantage of Wallerian degeneration to obtain more and activated SCs in culture comparing to intact nerve culture (Niapour et al 2010). The duration of 10 days has been implemented for predegeneration period in this research, based on other studies in which the 7-10 days in vitro predegeneration resulted in the highest SCs purity in primary cultures (Kraus et al 2010).…”

Section: Discussionmentioning

confidence: 99%

“…Our results indicated well adjustments of both SCs and FBs with this modification. Primary plated cells were cultured with 2.5 % KOSR since our previous work and other studies have demonstrated optimal concentration of serum allowing SCs proliferation and hindering of FBs expansion (Niapour et al 2010;Takagi et al 2011). Given the identity of SCs, the neural crest stem cells medium was considered and adapted to SCs culture through addition of chemically defined ingredients.…”

Section: Discussionmentioning

confidence: 99%

“…Once the epineurium stripped off under stereomicroscope, nerves were cut into short segments of 2-3 mm in length. Explants were incubated in predegeneration medium containing Dulbecco's modified eagles medium (DMEM, Invitrogen (Carlsbad, CA, SA), 12800-116) and 10 % fetal bovine serum (FBS, Invitrogen: 10270) supplemented with, 2 mM L-glutamine (Invitrogen: 25030), 100 U/ml penicillin/streptomycin (Invitrogen: 15140) for 10 days at 37°C and 5 % CO 2 (Niapour et al 2010). After predegeneration, explants were dissociated enzymatically for 20 h. Dissociation medium consisted of DMEM, 15 % FBS plus 0.125 % collagenase type IV (Invitrogen: 17104-019) and 1.25 U dispase (Invitrogen: 17105-041) per ml.…”

Section: Cultivation and Enrichment Of Scsmentioning

confidence: 99%

“…Yet, several techniques have been introduced and implemented in different laboratories in order to separate SCs from contaminating FBs. These modifications include: repeated explantations method (Morrissey et al 1995), combination of antimitotic treatment and antibody-mediated cytolysis which employs the use of complement (Assouline et al 1983;Brockes et al 1979), serum tapering (Komiyama et al 2003), manipulating differential adhesion and detachment proprieties of SCs and FBs (Jin et al 2008;Niapour et al 2010), immunoselective methods through fluorescent activated cell sorting (FACS) or magnetically activated cell sorting (MACS; Vroemen and Weidner 2003), and exploiting specific biochemical diversity between SCs and EBs (Kaewkhaw et al 2012). Requiring multiple steps, technically demanding procedures and utilizing animal products for culture are the main bottlenecks of reported systems.…”

Section: Introductionmentioning

confidence: 99%

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An efficient system for selection and culture of Schwann cells from adult rat peripheral nerves

et al. 2015

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Schwann cells (SCs), the supporting cells of the peripheral nerves, are indispensable for regenerating the peripheral and central nervous system. Copious preparation of these cells in a well-defined manner is to be a privileged position. SCs cultivation is overwhelmed by contaminating fibroblasts which are often outgrowing as the predominant cell type in an in vitro culture. This study introduces a technically simple and efficient procedure for SCs isolation and enrichment based on implementing recombinant and defined supplements.Collected adult rat sciatic nerves were cultured for 10 days as in vitro predegeneration. After dissociation and plating, the medium changed to knockout serum replacement supplemented DMDM/F12 medium containing various growth factors. The whole procedure took 3 weeks and SCs purity was then evaluated through implementing specific cytoplasmic and membranous markers. The viability of enriched SCs were evaluated by MTT assay. Within 10 days, over 99 % homogenous SCs were achieved and confirmed through immunofluorescence staining and flow-cytometry for P75 NTR and S100 markers, respectively. MTT data revealed that the viability and metabolic activities of purified SCs were increased in expansion medium. This study provides a technically easy and efficient method with the benefits of not utilizing bovine serum or other animal products for SCs isolation and enrichment.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Cultivation and Enrichment Of Scsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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An efficient system for selection and culture of Schwann cells from adult rat peripheral nerves

et al. 2015

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“…Although successful, such strategies are technically complicated, costly, and may be limited by the high number of cells required for each intervention, which carries the risk of potential injury to the donor nerve, donor-site morbidity, and fibroblast contamination of cultures (Niapour et al 2010). Therefore, alternative sources of cells have been sought.…”

Section: Nerve Guidance Conduits Supplemented With Autologous Schwannmentioning

confidence: 99%

Stem Cell-Based Approaches to Improve Nerve Regeneration: Potential Implications for Reconstructive Transplantation?

Khalifian

Sarhane

Tammia

et al. 2014

Arch. Immunol. Ther. Exp.

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Reconstructive transplantation has become a viable option to restore form and function after devastating tissue loss. Functional recovery is a key determinant of overall success and critically depends on the quality and pace of nerve regeneration. Several molecular and cell-based therapies have been postulated and tested in pre-clinical animal models to enhance nerve regeneration. Schwann cells remain the mainstay of research focus providing neurotrophic support and signaling cues for regenerating axons. Alternative cell sources such as mesenchymal stem cells and adipose-derived stromal cells have also been tested in pre-clinical animal models and in clinical trials due to their relative ease of harvest, rapid expansion in vitro, minimal immunogenicity, and capacity to integrate and survive within host tissues, thereby overcoming many of the challenges faced by culturing of human Schwann cells and nerve allografting. Induced pluripotent stem cell-derived Schwann cells are of particular interest since they can provide abundant, patient-specific autologous Schwann cells. The majority of experimental evidence on cell-based therapies, however, has been generated using stem cell-seeded nerve guides that were developed to enhance nerve regeneration across "gaps" in neural repair. Although primary end-to-end repair is the preferred method of neurorrhaphy in reconstructive transplantation, mechanistic studies elucidating the principles of cell-based therapies from nerve guidance conduits will form the foundation of further research employing stem cells in end-to-end repair of donor and recipient nerves. This review presents key components of nerve regeneration in reconstructive transplantation and highlights the pre-clinical studies that utilize stem cells to enhance nerve regeneration.

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Graphene Oxide Substrate Promotes Neurotrophic Factor Secretion and Survival of Human Schwann‐Like Adipose Mesenchymal Stromal Cells

et al. 2021

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Mesenchymal stromal cells from adipose tissue (AD‐MSCs) exhibit favorable clinical traits for autologous transplantation and can develop ‘Schwann‐like’ phenotypes (sAD‐MSCs) to improve peripheral nerve regeneration, where severe injuries yield insufficient recovery. However, sAD‐MSCs regress without biochemical stimulation and detach from conduits under unfavorable transplant conditions, negating their paracrine effects. Graphene‐derived materials support AD‐MSC attachment, regulating cell adhesion and function through physiochemistry and topography. Graphene oxide (GO) is a suitable substrate for human sAD‐MSCs incubation toward severe peripheral nerve injuries by evaluating transcriptome changes, neurotrophic factor expression over a 7‐days period, and cell viability in apoptotic conditions is reported. Transcriptome changes from GO incubation across four patients are minor compared to biological variance. Nerve growth factor (NGF), brain‐derived neurotrophic factor (BDNF), and glial‐derived neurotrophic factor (GDNF) gene expression is unchanged from sAD‐MSCs on GO substrates, but NGF and GDNF protein secretion increase at day 3 and 7. Secretome changes do not improve dorsal root ganglia neuron axon regeneration in conditioned media culture models. Fewer sAD‐MSCs detach from GO substrates compared to glass following phosphate buffer saline exposure, which simulates apoptotic conditions. Overall, GO substrates are compatible with sAD‐MSC primed for peripheral nerve regeneration strategies and protect the cell population in harsh environments.

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Novel method to obtain highly enriched cultures of adult rat Schwann cells

Cited by 28 publications

References 23 publications

An efficient system for selection and culture of Schwann cells from adult rat peripheral nerves

An efficient system for selection and culture of Schwann cells from adult rat peripheral nerves

Stem Cell-Based Approaches to Improve Nerve Regeneration: Potential Implications for Reconstructive Transplantation?

Graphene Oxide Substrate Promotes Neurotrophic Factor Secretion and Survival of Human Schwann‐Like Adipose Mesenchymal Stromal Cells

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