Selective expansion and long‐term culture of human schwann cells from sural nerve biopsies

Abstract

Fragments of sural nerve biopsy specimens were cultured in the presence of the supernatant of lymphokine-activated killer cells, resulting in the selective outgrowth of cells with bipolar or tripolar morphology, reminiscent of that of Schwann cells. Immunofluorescent staining with antibodies to the S-100 protein, the low-affinity nerve growth factor receptor, and the surface Thy-1 antigen confirmed that these cultures contained more than 99% Schwann cells and no detectable fibroblasts. The mitotic activity of Schwann cells was measured by bromodeoxyuridine labeling, and was increased when the cells were grown in medium with lymphokine-activated killer cell supernatant compared with medium without this supernatant. In the presence of lymphokine-activated killer cell supernatant, Schwann cells could be maintained in continuous culture for a minimum of 8 months.

Adhesion and proliferation of human Schwann cells on adhesive coatings

“…1 mm 3 . These were placed in gelatin-coated (see below) culture flasks (Greiner, The Netherlands) and covered with a thin layer of LAK culture medium [24], consisting of Dulbecco's modified Eagle's medium (DMEM; Bio-Whittaker Europe, Belgium), 10% lymphokine activated killer cells conditioned medium (LAK) [25], 5% fetal calf serum (FCS; Gibco BRL, Life Technologies, Germany), 0.25 ml/ml phytohaem-agglutinin (PHA; Difco Laboratories, USA), 100 IU/ml penicillin (Gist-Brocades, The Netherlands), and 50 mg/ml streptomycin (Gist-Brocades).…”

Mycobacterium leprae-Specific, HLA Class II-Restricted Killing of Human Schwann Cells by CD4+ Th1 Cells: A Novel Immunopathogenic Mechanism of Nerve Damage in Leprosy

“…Human Schwann cells were isolated from sural nerve biopsies from amputated material and propagated as described before (15). Briefly, sural nerve specimens were cut into small pieces and incubated in 85% IMDM, supplemented with 10% lymphokine-activated killer (LAK) cell supernatant, 5% FCS, 100 U/ml penicillin, 100 g/ml streptomycin, and 0.25 g/ml PHA (Murex Diagnostics, Dartford, U.K.), in humidified 5% CO 2 at 37°C.…”

Therapeutic strategies in multiple sclerosis. II. Long–term repair

“…Much is known from rodent studies of the behaviour and properties of Schwann cells, and of their interactions with other neural elements (see } 2(b)(ii)) (Ludwin 1988;Jessen & Mirsky 1997;Mirsky & Jessen 1999;Bunge 1987;Duncan & Ho¡man 1997;Reynolds & Woolf 1993), although (thus far) the biology of human Schwann cells has been less intensively explored; whether, as in the embryonic rodent (Jessen & Mirsky 1997 ), Schwann cell precursors are present in the human nervous system has not been ascertained. Importantly, however, there is good understanding of those growth factors which trigger Schwann cell proliferationöincluding that of human cells (Rutkowski et al 1995;Van den Berg et al 1995). Glial growth factors (neuregulins) are particularly important in this respect (Mirsky & Jessen 1999); they have also been implicated in the proliferation of adult rodent oligodendrocytes (Shi et al 1998).…”