Cold jet: a method to obtain pure Schwann cell cultures without the need for cytotoxic, apoptosis-inducing drug treatment

Jirsová, Kateřina; Sodaar, Peter; Mandys, Václav; Bär, P.R.

doi:10.1016/s0165-0270(97)00146-5

Cited by 44 publications

(43 citation statements)

References 24 publications

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“…The cell pellets obtained after centrifugation were then resuspended in Dulbecco's modified Eagle's medium (DMEM) containing 10% (vol/vol) heat-inactivated fetal bovine serum (FBS), plated at a density of 20,000 cells/cm 2 and grown for 2 days at 37 o C in a humidified atmosphere containing 5% CO2. After two or three subcultures using the coldjet method (Jirsova et al, 1997), the identity and purity of SCs were evaluated via immunostaining with an antibody raised against S100 (data not shown).…”

Section: Primary Schwann Cell Cultures and Rt4 Cellsmentioning

confidence: 99%

Extracellular Signal-regulated Kinase Activation Is Required for Serine 727 Phosphorylation of STAT3 in Schwann Cells in vitro and in vivo

Lee

Jung

Lee

et al. 2009

Korean J Physiol Pharmacol

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In the peripheral nerves, injury-induced cytokines and growth factors perform critical functions in the activation of both the MEK/ERK and JAK/STAT3 pathways. In this study, we determined that nerve injury-induced ERK activation was temporally correlated with STAT3 phosphorylation at the serine 727 residue. In cultured Schwann cells, we noted that ERK activation is required for the serine phosphorylation of STAT3 by neuropoietic cytokine interleukin-6 (IL-6). Serine phosphorylated STAT3 by IL-6 was transported into Schwann cell nuclei, thereby indicating that ERK may regulate the transcriptional activity of STAT3 via the induction of serine phosphorylation of STAT3. Neuregulin-1 (NRG) also induced the serine phosphorylation of STAT3 in an ERK-dependent fashion. In contrast with the IL-6 response, serine phosphorylated STAT3 induced by NRG was not detected in the nucleus, thus indicating the non-nuclear function of serine phosphorylated STAT3 in response to NRG. Finally, we determined that the inhibition of ERK prevented injury-induced serine phosphorylation of STAT3 in an ex-vivo explants culture of the sciatic nerves. Collectively, the results of this study show that ERK may be an upstream kinase for the serine phosphorylation of STAT3 induced by multiple stimuli in Schwann cells after peripheral nerve injury.

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Section: Primary Schwann Cell Cultures and Rt4 Cellsmentioning

confidence: 99%

Extracellular Signal-regulated Kinase Activation Is Required for Serine 727 Phosphorylation of STAT3 in Schwann Cells in vitro and in vivo

Lee

Jung

Lee

et al. 2009

Korean J Physiol Pharmacol

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“…We have purified and characterised Schwann cells using a combination of various published methods (Jirsova et al, 1997;Komiyama et al, 2003;Shen et al, 2002;Tanaka and Webster, 1993;Watabe et Fig. 4.…”

Section: Discussionmentioning

confidence: 99%

“…Although the detailed mechanism is still unknown, it is likely that the spontaneous immortalisation of long term cultured Schwann cells is a general phenomenon in both wild type and mutated mice (Watabe et al, 2003). To date more than 15 Schwann cell lines have been established by spontaneous immortalization or transfection with oncogenes (Bolin et al, 1992;Boutry et al, 1992;Chen et al, 1987;Goda et al, 1991;Jirsova et al, 1997;Li et al, 1996;Ridley et al, 1988;Tennekoon et al, 1987;Thi et al, 1998;Watabe et al, 1990Watabe et al, , 1995. The degree of phenotype expression and differentiation differ from each other (Hai et al, 2002).…”

Section: Discussionmentioning

confidence: 99%

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A spontaneously immortalized Schwann cell line to study the molecular aspects of metachromatic leukodystrophy

Saravanan

Büssow

Weiler

et al. 2007

Journal of Neuroscience Methods

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“…The abundant connective tissues in nerves further complicate their enrichment during the following purification procedure. To date, several methods have been described for obtaining SCs, including antimitotic treatment (Calderon-Martinez et al 2002), a combination of antimitotic treatment and antibody-mediated cytolysis by employing complements (Brockes et al 1979), a repeated explantation method (Oda et al 1989), differential adhesion methods (Pannuzio et al 2005), immunoselective methods (Manent et al 2003), in vivo or in vitro predegeneration (Komiyama et al 2003;Mauritz et al 2004), the cold jet technique (Jirsová et al 1997), differential detachment methods (Jin et al 2008), and a combination of the in vitro predegeneration and cold jet technique (Haastert et al 2007(Haastert et al , 2009). These methods can provide highly purified SCs with various efficiencies.…”

Section: Introductionmentioning

confidence: 99%

An improved method for isolating Schwann cells from postnatal rat sciatic nerves

et al. 2009

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The major difficulty in Schwann cell (SC) purification is contamination by fibroblasts, which usually become the predominant cell type during SC enrichment in vitro. Current reported measures are mainly limited by either high cost or complicated procedures with low cell yields or purity. Our objectives have been to develop an efficient, easily applicable, rapid method to obtain highly purified SC from the sciatic nerve of newborn rats. The method involves two rounds of purification to eliminate fibroblasts with the novel combined use of cytosine-B-arabinoside hydrochloride (Ara-C) action and differential cell detachment. Cultured cells were first treated with Ara-C for 24 h. The medium was replaced with the growth medium containing 20 ng/ml human heregulin1-beta1 extracellular domain (HRG1-beta1 ECD). After another 48 h in culture, the cells were treated with 0.05% trypsin, following which SCs, but not fibroblasts, were easily detached from the dishes. The advantage of this method is that the two steps can eliminate the fibroblasts complementarily. Ara-C eliminates most of the fibroblasts growing among SCs, whereas the differential cell detachment technique removes the remainder growing under or interacting with the SC layer. A purity of more than 99% SCs has been obtained, as confirmed by cell morphology and immunostaining. The purified SCs have a spindle-shaped, bipolar, and sometimes tripolar morphology, align in fascicles, and express S-100. The whole procedure takes about 10 days from primary culture to the purified SCs growing to confluence (only half the time reported previously). This protocol provides an alternative method for investigating peripheral nerve regeneration and potentially could be used to produce enough SCs to construct artificial nerve scaffolds in vitro.

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Cold jet: a method to obtain pure Schwann cell cultures without the need for cytotoxic, apoptosis-inducing drug treatment

Cited by 44 publications

References 24 publications

Extracellular Signal-regulated Kinase Activation Is Required for Serine 727 Phosphorylation of STAT3 in Schwann Cells in vitro and in vivo

Extracellular Signal-regulated Kinase Activation Is Required for Serine 727 Phosphorylation of STAT3 in Schwann Cells in vitro and in vivo

A spontaneously immortalized Schwann cell line to study the molecular aspects of metachromatic leukodystrophy

An improved method for isolating Schwann cells from postnatal rat sciatic nerves

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