Separation of functional Schwann cells and neurons from normal peripheral nerve tissue

Wood, Patrick M.

doi:10.1016/0006-8993(76)90355-3

Cited by 337 publications

(180 citation statements)

References 18 publications

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“…Cultures were incubated at 37°C with a 95% air/5% CO 2 gas mix in Eagle's minimum essential medium with glutamine supplemented with 10% glucose, 5% fetal bovine serum (FBS) and 0.01% nerve growth factor (2.5). The medium was changed every two days, alternating between the above medium and medium containing the antimitotic agent, 5-fluorodeoxyuridine (10 −5 M; Sigma-Aldrich), which limits the proliferation of Schwann cells in DRG cultures (Wood, 1976). Following a 10-day incubation, the cultures were fixed in 4% paraformaldehyde in 0.1 M phosphate-buffered saline (PBS).…”

Section: Drg Isolation and Culturementioning

confidence: 99%

Evaluating neuronal and glial growth on electrospun polarized matrices: bridging the gap in percussive spinal cord injuries

et al. 2007

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One of the many obstacles to spinal cord repair following trauma is the formation of a cyst that impedes axonal regeneration. Accordingly, we examined the potential use of electrospinning to engineer an implantable polarized matrix for axonal guidance. Polydioxanone, a resorbable material, was electrospun to fabricate matrices possessing either aligned or randomly oriented fibers. To assess the extent to which fiber alignment influences directional neuritic outgrowth, rat dorsal root ganglia (DRGs) were cultured on these matrices for 10 days. Using confocal microscopy, neurites displayed a directional growth that mimicked the fiber alignment of the underlying matrix. Because these matrices are generated from a material that degrades with time, we next determined whether a glial substrate might provide a more stable interface between the resorbable matrix and the outgrowing axons. Astrocytes seeded onto either aligned or random matrices displayed a directional growth pattern similar to that of the underlying matrix. Moreover, these glia-seeded matrices, once cocultured with DRGs, conferred the matrix alignment to and enhanced outgrowth exuberance of the extending neurites. These experiments demonstrate the potential for electrospinning to generate an aligned matrix that influences both the directionality and growth dynamics of DRG neurites.

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Section: Drg Isolation and Culturementioning

confidence: 99%

Evaluating neuronal and glial growth on electrospun polarized matrices: bridging the gap in percussive spinal cord injuries

et al. 2007

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“…Cultures were prepared in 25 mm Aclar fluorocarbon dishes or on 22 mm acid-cleaned glass coverslips, as described previously (Masurovsky and Bunge, 1968;Wood, 1976;Johnson and Argiro, 1983). Collagen substrata were prepared from acetic acid-extracted rat tail collagen either by air-drying at room temperature (ADC) or by a 2 min exposure to ammonia vapors, rinsing with water, and air-drying (ammoniated collagen) (Wood, 1976;Johnson and Argiro, 1983). Laminin (Collaborative Research; 5 pg in 30 ~1 of 0.05 M carbonate buffer, pH 9.6) was applied to 22 mm glass coverslips at 35°C overnight.…”

Section: Methodsmentioning

confidence: 99%

Schwann cell surfaces but not extracellular matrix organized by Schwann cells support neurite outgrowth from embryonic rat retina

et al. 1988

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Despite evidence that glial cell surfaces and components of the extracellular matrix (ECM) support neurite outgrowth in many culture systems, the relative contributions of these factors have rarely been compared directly. Specifically, it remains to be determined which components of peripheral nerve support growth of central nerve fibers. We have directly compared neurite outgrowth from embryonic day 15 rat retinal explants placed onto beds of (1) Schwann cells without ECM, (2) Schwann cells expressing ECM (including a basal lamina), (3) cell-free ECM prepared from neuron-Schwann cell cultures, (4) nonglial cells (fibroblasts), and (5) 2 isolated ECM components, laminin and type I collagen. From the first day in culture, retinal explants extended neurites when placed on Schwann cells without ECM. Outgrowth on Schwann cells expressing ECM was also extensive, but not obviously different form that on Schwann cells alone. Ultrastructural study revealed that 95% of retinal neurites in ECM-containing cultures contacted other neurites and Schwann cell surfaces exclusively. On cell-free ECM prepared from neuron-Schwann cell cultures, neurite extension was poor to nonexistent. No neurite outgrowth occurred on fibroblasts. Retinal explants also failed to extend neurites onto purified laminin and ammoniated type I collagen substrata; however, growth was rapid and extensive on air-dried type I collagen. In cultures containing islands of air-dried type I collagen on a laminin-coated coverslip, retinal explants attached and extended neurites on collagen, but these neurites did not extend off the island onto the laminin substratum. We conclude from these experiments that neurite extension from embryonic rat retina is supported by a factor found on the surface of Schwann cells and that neither organized nor isolated ECM components provide this neurite promotion. These findings are discussed in relation to possible species differences in growth requirements for retinal ganglion cell neurites and to the specificity of response of different CNS neurites to ECM substrata.

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“…When mitoses cease, SC quickly recover their SC phenotype with rhythmic undulations while migration speed increased to 92±20 µ/h. Thus, in spite of dramatic modification of shape, structure and behavior during mitotic stimulation, SC subsequently recover their unique motility pattern which might be essential for their myelinating function.Rat Schwann cells (SC), the myelin-form-ing cells of the peripheral nervous system, can now be cultured in isolation by using two different techniques [1][2][3]. In the first, rat dorsal root ganglia obtained just before birth are explanted and treated with antimitotic agents for a few days.…”

mentioning

confidence: 99%

“…Rat Schwann cells (SC), the myelin-form-ing cells of the peripheral nervous system, can now be cultured in isolation by using two different techniques [1][2][3]. In the first, rat dorsal root ganglia obtained just before birth are explanted and treated with antimitotic agents for a few days.…”

mentioning

confidence: 99%

“…In the first, rat dorsal root ganglia obtained just before birth are explanted and treated with antimitotic agents for a few days. After 10 days, the ganglia are excised and a "Schwann cell bed" is developed after the neurites are eliminated [1,2]. In the second technique, dissociation of newborn rat sciatic nerve results in a mixed population of SC and fibroblasts, which after 2 days of culturing are identified by their respective surface antigenic markers Ran 1 and Thy 1.1 [3,4].…”

mentioning

confidence: 99%

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Structure and behavior of rat primary and secondary Schwann cells in vitro

Dubois‐Dalcq

Rentier

et al. 1981

Experimental Cell Research

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SUMMARYThe structure and motility of isolated rat primary (I) Schwann cells (SC) have been compared to that of subcultured (II) SC during and after mitotic stimulation. I SC contain myelin components which persist for 2 weeks in serum-free medium while they rapidly disappear in medium containing serum and high glucose concentration. These components were never detected in II SC. Both I SC and II SC after their mitotic phase are spindle-shaped, contain many intermediate and actin filaments, have no basement membrane but show intense migratory and undulatory activities. Rare fibroblasts in I cultures are recognized by their extremely variable shape, the presence of Thy 1.1 antigen in their membrane and their intense edge ruffling alternating with abrupt translocation. In contrast, I SC movements consist of intracellular translocation of nuclei along SC processes, which retract and extend constantly, and in slow rhythmic undulation episodes (2.3±0.2/min) alternating with migration at 135±50 µ/h. The total number of these episodes per day in serum-free medium is rigorously identical for different cells (166.3±0.2) and this uniformity of frequency suggests a genotypic basis. Cycles, consisting of an undulation episode followed by a resting interval, have mean durations of 8.6±4.1 min and a sharp peak of occurrence at 6 min, with exponential distribution of the longer periods. Motility of II SC is considerably inhibited during mitotic stimulation by cholera toxin and a pituitary extract while SC phenotype has changed to a flat multipolar cell with prominent Golgi and ribosomes. Migration is reduced to 24±2 µ/h and only 2 % of the SC show pulsations of the same periodicity as the I SC undulations. A dramatic increase in pulsation frequency occurs 6-12 h after removal of mitogenic factors when 80% of II SC start pulsating twice as fast for 2-3 days. When mitoses cease, SC quickly recover their SC phenotype with rhythmic undulations while migration speed increased to 92±20 µ/h. Thus, in spite of dramatic modification of shape, structure and behavior during mitotic stimulation, SC subsequently recover their unique motility pattern which might be essential for their myelinating function.Rat Schwann cells (SC), the myelin-form-ing cells of the peripheral nervous system, can now be cultured in isolation by using two different techniques [1][2][3]. In the first, rat dorsal root ganglia obtained just before birth are explanted and treated with antimitotic agents for a few days. After 10 days, the ganglia are excised and a "Schwann cell bed" is developed after the neurites are eliminated [1,2]. In the second technique, dissociation of newborn rat sciatic nerve results in a mixed population of SC and fibroblasts, which after 2 days of culturing are identified by their respective surface antigenic markers Ran 1 and Thy 1.1 [3,4]. In these cultures, fibroblast growth is first inhibited by AraC treatment and subsequently, by complement-mediated killing of Thy 1.1 positive cells [3]. SC growth can then be intensely stimulated by pi...

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Separation of functional Schwann cells and neurons from normal peripheral nerve tissue

Cited by 337 publications

References 18 publications

Evaluating neuronal and glial growth on electrospun polarized matrices: bridging the gap in percussive spinal cord injuries

Evaluating neuronal and glial growth on electrospun polarized matrices: bridging the gap in percussive spinal cord injuries

Schwann cell surfaces but not extracellular matrix organized by Schwann cells support neurite outgrowth from embryonic rat retina

Structure and behavior of rat primary and secondary Schwann cells in vitro

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