Base media (plating medium and co-culture medium) were prepared as previously described [25 (link)]. Briefly, plating medium consisted of Neurobasal A culture medium supplemented with 2% B27 supplement, 1x Glutamax, 10% heat-inactivated horse serum, and 1 M HEPES at pH 7.5, while the co-culture medium consisted of Neurobasal A culture medium supplemented with 2% B27 supplement and 1x Glutamax (all from ThermoFisher). The tri-culture medium consisted of supplementing the co-culture medium with 100 ng/mL mouse IL-34 (R&D Systems), 2 ng/mL TGF-β (Peprotech), and 1.5 μg/mL ovine wool cholesterol (Avanti Polar Lipids). Due to the limited shelf life of IL-34 and TGF-β, the tri-culture medium was made fresh each week.
Mouse il 34
Manufactured by R&D Systems
Mouse IL-34 is a recombinant mouse protein that belongs to the Interleukin-34 family. It is a cytokine that promotes the survival and differentiation of monocytes and macrophages.
Automatically generated - may contain errors
2 protocols using mouse il 34
1
Tri-culture microglia-neuron-astrocyte protocol
2
Primary Neuron Tri-Culture Protocol
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Base media (plating medium and co-culture medium) were prepared as previously described (Chapman et al., 2015) (link). Briefly, plating medium consisted of Neurobasal A culture medium supplemented with 2% B27 supplement, 1x Glutamax, 10% heat-inactivated horse serum and 1 M HEPES at pH 7.5, while the co-culture medium consisted of Neurobasal A culture medium supplemented with 2% B27 supplement and 1x Glutamax (all from ThermoFisher). The tri-culture medium consisted of supplementing the co-culture medium with 100 ng/mL mouse IL-34 (R&D Systems), 2 ng/mL TGF-β (Peprotech) and 1.5 µg/mL ovine wool cholesterol (Avanti Polar Lipids). Due to the limited shelf life of the IL-24 and TGF-β, tri-culture medium was made new each week.
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