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

Abstract

Schwann cells (SCs) are indispensable for cell therapy and tissue engineering of the peripheral nervous system. Easy access to activated, highly proliferative SCs is necessary for clinical applications. The present study developed a fast, efficient method for obtaining highly purified SCs from the peripheral nerve of a mature Rhesus monkey. The common peroneal nerves of 4-year-old Rhesus monkeys were harvested and subjected to in vitro pre-degeneration in a modified SC culture medium (SCCM). The nerve pieces were subsequently treated enzymatically to dissociate the cells and then cultured for 2 days in SCCM. Cultured cells were treated with purification medium containing Ara-C to assist in restricting the overgrowth of fibroblast-like cells, for 24 h. After another 24-h cultivation period, the cells were subsequently treated with a multiplex collagenase, which enabled SC detachment over fibroblast detachment, and thereby facilitated SC isolation. Finally, SCs were cultured in SCCM. The cell yield was determined by cell counting following enzyme digestion and SC purity was determined from the percentage of SCs with respect to the total number of cells. Following purification, 96.3±3.9% of cells were identified as SCs. In vitro pre-degeneration in the presence of basic-fibroblast growth factor, heregulin β1 and forskolin maximized the purity and yield of SCs that could be obtained from monkey peroneal nerves. The present study identified a novel technique that can efficiently isolate and purify SCs from mature monkey nerves based on in vitro pre-degeneration.

Identification of adhesion‑associated DNA methylation patterns in the peripheral nervous system

“…Proliferation, adhesion and migration may markedly change with the epigenetic changes that occur in ASCs. Therefore, Schwann cell transplantation in peripheral nerve repair requires to overcome numerous difficulties ( 71 ). The present study performed bioinformatics analyses of DNA methylation patterns associated with Schwann cell adhesion and proliferation.…”

Dataset on inflammation induced after lumbar puncture

“…The samples were loaded in the wells with 12% gels (Bio-Rad Laboratories, CA, USA) and transferred to the membrane (PVDF, Immobilon-P, Millipore, Billerica, MA, USA). The membranes were blocked in 5% Milk in Tris-buffered saline Tween-20 (TBST) solution for 1h at room temperature and incubated overnight at 4 °C with the following primary antibodies: rabbit anti-Iba1 (1:1000, Wako 019-19741), rabbit anti-GFAP (1:1000, #AB7779 Abcam), rabbit anti-S100 (1:1000, Z0311, Dako, strongly reacts with S100B [8] , weakly with A1 and very weakly with A6) and rabbit anti-beta actin (1:100000, #A3854 Sigma). The membranes were then incubated with the following secondary antibody: HRP donkey conjugated-anti-rabbit (1:10000, #31430 Thermo Scientific).…”

“…Unexpected neuroinflammation has occurred 2-3 weeks after performing the given procedure. Data presented in the article implicate active microglia measured by using protein biomarker such as Iba-1 [1 , 3] and astrocytes measured by using Glial fibrillary acidic protein (GFAP) [2 , 3] and S100 (strongly targets S100B and weakly A1) [7 , 8] in the CSF collected two-three weeks after the laminectomy and dura breakage from the adult rats. Later paraformaldehyde (PFA) fixed spinal cord slices were also collected from the animals and immunolabelled with the same biomarkers.…”

Synergetic Action of Forskolin and Mevastatin Induce Normalization of Lipids Profile in Dyslipidemic Rats through Adenosine Monophosphate Kinase Upregulation

“…Forskolin, the new/old compound that extracted from plant Coleus forskohlii , is used widely to stimulate adenylate cyclase (AC) that lead to elevate intracellular cAMP levels [ 9 , 10 ]. So it has been used for controlling many disorders like glaucoma [ 11 ], nerve impairments [ 12 ], and improve nerve cells [ 13 ], and blood pressure [ 14 ]. As well as it has been used in many studies for controlling of hyperlipidemias [ 15 , 16 ].…”