Daisuke Noto and Hiroshi Sakuma contributed equally to this work.
Development of a culture system to induce microglia-like cells from haematopoietic cells
Version of Record online: 25 SEP 2014
© 2013 The Authors. Neuropathology and Applied Neurobiology published by John Wiley & Sons Ltd on behalf of the British Neuropathological Society.
This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
Neuropathology and Applied Neurobiology
Volume 40, Issue 6, pages 697–713, October 2014
How to Cite
Noto, D., Sakuma, H., Takahashi, K., Saika, R., Saga, R., Yamada, M., Yamamura, T. and Miyake, S. (2014), Development of a culture system to induce microglia-like cells from haematopoietic cells. Neuropathology and Applied Neurobiology, 40: 697–713. doi: 10.1111/nan.12086
Authors have no conflict of interest to declare.
- Issue online: 25 SEP 2014
- Version of Record online: 25 SEP 2014
- Accepted manuscript online: 10 SEP 2013 03:32AM EST
- Manuscript Accepted: 2 SEP 2013
- Manuscript Received: 23 MAY 2013
- Japan Health Sciences Foundation. Grant Number: KHB1010
Figure S1. Effect of human IL-34 on the differentiation of human ML cells. (A) Fluorescent microscopic images of human monocytes cocultured with human astrocytes in the presence or absence of M-CSF (50 ng/ml) or IL-34 (50 ng/ml). Data are representative of three independent experiments. Bar: 100 μm. (B) Dose–response relationship between M-CSF/IL-34 and the number of ML cells differentiated from monocytes (cell number per mm2). Data are presented as mean ± SD. *P < 0.05 as compared with untreated cells. (C) Immunofluorescence of human ML cells. Human monocytes culture with human IL-34 (50 ng/ml) mainly showed spindle-like morphology (arrow) and all cells were positive for TREM2. Data are representative of three independent experiments. Bar: 50 μm.
Figure S2. Functional characterization of isolated ML cells. (A) Morphology of peritoneal macrophages, microglia isolated from mixed glial culture, and murine ML cells isolated from astrocyte coculture. Data are representative of three independent experiments. Bar: 50 μm. (B) Immunofluorescence of isolated murine ML cells. All cells were positively stained with Iba1, CD11b, TREM2 and MHC II. Data are representative of three independent experiments. Bar: 100 μm. (C) Phagocytosis assay: percentage of peritoneal macrophages, microglia and murine ML cells phagocytosing zymosan-coated particles. Data are presented as mean ± SD. (D) Cytokine production after LPS treatment. Cells were treated with or without LPS (20 ng/ml) and concentration of TNFα and IL-6 in culture supernatants were determined by ELISA. Incubation periods were 6 h for TNFα and 24 h for IL-6. Data are presented as mean ± standard error (SE).
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