Schwann cell-enriched cultures from adult human peripheral nerve: a technique combining short enzymatic dissociation and treatment with cytosine arabinoside (Ara-C)

Calderón-Martı́nez, Diany; Garavito, Zayra; Spinel, Clara; Hurtado, Hernán

doi:10.1016/s0165-0270(01)00493-9

Cited by 36 publications

(33 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The pre-degeneration approach results in a higher yield of SCs relative to the intact nerve approach (Keilhoff et al 2000;Mauritz et al 2004). However, to obtain purified and homogenous SCs different modifications to the pre-degeneration approach have been applied which include: (i) antimitotic treatment such as the arabinoside chlora toxin (Calderon-Martinez et al 2002), (ii) a combination of antimitotic treatment and antibody-mediated cytolysis which employs the use of complements (Assouline et al 1983), (iii) repeated explantation methods (Oda et al 1989;Morrissey et al 1995a, b), (iv) differential adhesion and detachment methods (Mauritz et al 2004;Pannunzio et al 2005;Jin et al 2008), (v) immunoselective methods (Manent et al 2003;Vroemen and Weidner 2003), and (vi) serum tapering (Komiyama et al 2003;Hedayatpour et al 2007).…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2010

View full text Add to dashboard Cite

Schwann cells (SCs) can be used to repair both the peripheral and central nervous systems. Therefore, establishment of a procedure to obtain activated, highly proliferative SCs, in an appropriate time for clinical applications, is a prerequisite. Purification is complicated by contamination with fibroblasts which often become the predominant cell type in an in vitro SC culture. This study describes a novel and efficient method to enrich SCs by utilizing the differential detachment properties of the two cell types. In culture, cells were treated with two different media and the chelator, EGTA, which detached SCs faster than fibroblasts and allowed for easy isolation of SCs. Within seven days, high yields of SCs with a purity of greater than 99% were achieved. This was confirmed by immunostaining characterization and flow-cytometric analyses using an antibody against the p75 low affinity nerve growth factor receptor (p75LNGFR). The entire procedure was completed in approximately 21 days. This method has the advantage of being technically easier, faster, and more efficient than other previously described methods. An SC culture that was about 99% homogenous was achieved.

show abstract

Section: Introductionmentioning

confidence: 99%

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

et al. 2010

View full text Add to dashboard Cite

show abstract

“…One practical difficulty with the potential use of Schwann cells for autologoues transplantation is that autologous Schwann cells will not be available immediately after a nerve lesion. The generation of sufficient quantities of Schwann cells for transplantation from the patient's own peripheral nerve biopsy requires at least 3-6 weeks according to established protocols (Morrissey et al, 1991;Calderon-Martinez et al, 2002). And, one or more functioning nerves must be sacrificed with the consequence of loss of sensation, scarring, and possibly, neuroma formation.…”

Section: Introductionmentioning

confidence: 99%

Transdifferentiated Mesenchymal Stem Cells as Alternative Therapy in Supporting Nerve Regeneration and Myelination

et al. 2006

View full text Add to dashboard Cite

show abstract

“…In order to prepare the SC-CM, Schwann cells were first isolated using a method similar to the one published by Calderon-Martinez et al (2002), using human peripheral nerves. In the present case, pairs of adult rat sciatic nerves were rapidly excised from animals sacrificed with CO 2 and once the epineurium was removed, nerve fascicles were cut in thin slices and cultured in DMEM with 10% FBS for 14 days, changing the medium every 3 days.…”

Section: Schwann Cell Conditioned Medium Sc-cmmentioning

confidence: 99%

Neuron-Like Differentiation of PC12 Cells Treated With Media Conditioned by Either Sciatic Nerves, Optic Nerves, or Schwann Cells

Villegas

Núñez

et al. 2005

Cell Mol Neurobiol

View full text Add to dashboard Cite

In previous works we reported the finding of neurotrophic activity in a serum-free Dulbecco's modified Eagle's medium conditioned by rat sciatic nerves, previously maintained in culture for 11 days. This medium produces rapid neuron-like differentiation of cultured PC12 cells, as revealed by an increase in the size of the cell body and by the extension of short and/or long neurites by most of the cells. Neuregulin present in the conditioned medium was demonstrated to play a key role in the observed differentiation. In the present work, taking into consideration those latter results, the neurotrophic activity of conditioned media prepared with sciatic and optic nerves cultured during days 1-4 and 9-12 were studied. Evaluation of the trophic activities of those media revealed an opposite timing in the activities of sciatic and optic nerves conditioned media. The activity of the sciatic nerve was not observed in the 1-4-day period, increasing then up to the 9-12-day period. On the contrary, the optic nerve conditioned medium was active in the 1-4-day period, decreasing down to the 9-12-day period. These results led us to explore the contribution of the different cellular constituents of those nerves to their neurotrophic properties. As a first step in that direction we also investigated the neurotrophic activity of media conditioned during 12 days by cultured Schwann cells isolated from rat sciatic nerves. The Schwann cell conditioned media did produce a rapid differentiation of the PC12 cells similar to that caused by the sciatic nerve conditioned medium, though of a lower magnitude. Variations in the trophic activities of the conditioned media used in the present work is discussed taking into consideration the production of trophic and inhibitory factors by the peripheral and central glial cells. The role played by the optic nerve glia and myelin is being investigated at present.

show abstract

Schwann cell-enriched cultures from adult human peripheral nerve: a technique combining short enzymatic dissociation and treatment with cytosine arabinoside (Ara-C)

Cited by 36 publications

References 20 publications

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

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

Transdifferentiated Mesenchymal Stem Cells as Alternative Therapy in Supporting Nerve Regeneration and Myelination

Neuron-Like Differentiation of PC12 Cells Treated With Media Conditioned by Either Sciatic Nerves, Optic Nerves, or Schwann Cells

Contact Info

Product

Resources

About