Isolation and Purification of Schwann Cells

SCs were isolated as previously described (39 (link), 40 (link)). In short, sciatic nerves were digested with dispase II (Sigma-Aldrich) and collagenase type I (Sigma-Aldrich) overnight. The resulting single-cell suspension was incubated for 5 d in a solution containing 2 µM forskolin (Sigma-Aldrich) and 10 nM neuregulin (PeproTech) to promote SC differentiation. After the separation of fibroblasts by magnetic cell selection, an SC purity above 95% was achieved. SC purity was controlled by FACS analysis with the SC marker SOX10 (1:1,000, rabbit recombinant monoclonal antibody, ab155279, RRID:AB_2650603, Abcam) after selection of viable cells (fixable viability dye eFlour 780, Thermo Fisher Scientific) as described by the manufacturer’s protocol.

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Grüter T., Mohamad N., Rilke N., Blusch A., Sgodzai M., Demir S., Pedreiturria X., Lemhoefer K., Gisevius B., Haghikia A., Fisse A.L., Motte J., Gold R, & Pitarokoili K. (2023). Propionate exerts neuroprotective and neuroregenerative effects in the peripheral nervous system. Proceedings of the National Academy of Sciences of the United States of America, 120(4), e2216941120.

Publication 2023

dispase II collagenase type I forskolin neuregulin ab155279 eFlour 780

Cells Collagenase i Dispase ii Fibroblasts Forskolin Monoclonal antibody Neuregulin Rabbit Sciatic nerves Sox10

Corresponding Organization :

Other organizations : Ruhr University Bochum, St. Josef-Hospital, Otto-von-Guericke University Magdeburg

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Variable analysis

independent variables

Digestion of sciatic nerves with dispase II and collagenase type I overnight
Incubation of the resulting single-cell suspension for 5 days in a solution containing 2 μM forskolin and 10 nM neuregulin

dependent variables

SC purity above 95% after the separation of fibroblasts by magnetic cell selection
SC purity controlled by FACS analysis with the SC marker SOX10

control variables

Viable cells selected for FACS analysis using fixable viability dye eFlour 780

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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