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

Abstract

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.

Combination treatment of all-trans retinoic acid (ATRA) and γ-secretase inhibitor (DAPT) cause growth inhibition and apoptosis induction in the human gastric cancer cell line

“…MTT assay was performed as described elsewhere (Niapour et al 2016a;Sharifi Pasandi et al 2017). Briefly, at the end of the treatment, 20 ll of MTT solution (5 mg/ml in PBS) was added to 180 ll medium in each well and plates were incubated in the dark for 4 h at 37°C.…”

Advances and Perspectives in Dental Pulp Stem Cell Based Neuroregeneration Therapies

“…Owing to the intimate relationship of hDPSCs with the peripheral nervous system, it is not surprising that they can in vitro differentiate to functional Schwann cells. A high proportion of hDPSCs exposed to neural differentiation media co-express p75NTR with S-100β [ 71 ], which constitutes the distinctive molecular marker profile of Schwann cells [ 97 ]. In vitro hDPSC-derived Schwann cells have been shown to help guide axonal extension and myelination [ 98 , 99 ].…”

A novel method for obtaining highly enriched Schwann cell populations from mature monkey nerves based on in vitro pre-degeneration

“…Experimentally, embryonic or newborn mice and rats are currently the predominant source of non-myelinating SCs, which have a strong capacity to proliferate ( 23 ). However, in most treatment strategies, SCs are obtained from adult animals.…”