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Updated: Sep 27, 2025
Primary Cultures of Rat Astrocytes and Microglia and Their Use in the Study of Amyotrophic Lateral Sclerosis
Published on: June 23, 2022
Bone marrow-derived inducible microglia-like cells ameliorate motor function and survival in a mouse model of
Shuhei Kobashi1, Tomoya Terashima2, Miwako Katagi2
1Department of Stem Cell Biology and Regenerative Medicine, Shiga University of Medical Science, Otsu, Japan; Department of Neurology, Shiga University of Medical Science, Otsu, Japan.
Background Aims:
Amyotrophic lateral sclerosis (ALS) is an incurable neurodegenerative disease. Neuroinflammation in the spinal cord plays a pivotal role in the pathogenesis of ALS, and microglia are involved in neuroinflammation. Microglia mainly have two opposite phenotypes involving cytotoxic and neuroprotective properties, and neuroprotective microglia are expected to be a novel application for the treatment of ALS. Therefore, to establish a clinically applicable therapeutic method using neuroprotective microglia, the authors investigated the effect of inducing neuroprotective microglia-like cells from bone marrow for transplantation into ALS model mice.
Methods:
Bone marrow-derived mononuclear cells were isolated from green fluorescent protein mice and cultured using different protocols of cytokine treatment with granulocyte-macrophage colony-stimulating factor (GM-CSF) and IL-4. Cells with a high potency of proliferation and differentiation into microglia were evaluated by gene analysis, flow cytometry and direct neuroprotective effects in vitro. These cells were named bone marrow-derived inducible microglia-like (BM-iMG) cells and transplanted into the spinal cords of ALS model mice, and behavioral tests, immunohistochemistry and gene expression profiling were performed.
Results:
Three-day GM-CSF and 4-day GM-CSF + IL-4 stimulations were most effective in inducing BM-iMG cells from the bone marrow. Transplantation of BM-iMG cells improved motor function, prolonged survival and suppressed neuronal cell death, astrogliosis and microgliosis in the spinal cords of ALS mice. Moreover, neuroprotective genes such as Arg1 and Mrc1 were upregulated, whereas pro-inflammatory genes such as Nos2 and Il6 were downregulated.
Conclusions:
Intraspinal transplantation of BM-iMG cells demonstrated therapeutic effects in a mouse model of ALS. Further studies and clinical applications in patients with ALS are expected in the future.
Insights
Transplanting bone marrow-derived inducible microglia-like (BM-iMG) cells into ALS model mice improved motor function and survival. This study shows potential for BM-iMG cells as a novel therapy for amyotrophic lateral sclerosis (ALS).
Area of Science:
- Neuroscience
- Immunology
- Regenerative Medicine
Background:
- Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease where spinal cord neuroinflammation, involving microglia, is key.
- Microglia exhibit both damaging and protective functions; neuroprotective microglia offer a potential therapeutic avenue for ALS.
- Developing a clinical method using neuroprotective microglia requires inducing these cells from a readily available source.
Purpose of the Study:
- To investigate the therapeutic potential of transplanting bone marrow-derived inducible microglia-like (BM-iMG) cells into a mouse model of ALS.
- To establish a method for generating neuroprotective microglia-like cells from bone marrow for potential ALS treatment.
- To evaluate the effects of BM-iMG cell transplantation on motor function, survival, and neuropathology in ALS mice.
Main Methods:
- Bone marrow mononuclear cells from GFP mice were cultured with GM-CSF and IL-4 to induce BM-iMG cells.
- BM-iMG cell potency was assessed via proliferation, differentiation, gene analysis, flow cytometry, and in vitro neuroprotection assays.
- BM-iMG cells were transplanted into the spinal cords of ALS model mice, followed by behavioral, histological, and gene expression analyses.
Main Results:
- Optimal induction of BM-iMG cells was achieved with 3-day GM-CSF and 4-day GM-CSF + IL-4 stimulation.
- Transplantation of BM-iMG cells significantly improved motor function and prolonged survival in ALS mice.
- BM-iMG cell therapy reduced neuronal death, astrogliosis, and microgliosis, upregulating neuroprotective genes (Arg1, Mrc1) and downregulating inflammatory genes (Nos2, Il6).
Conclusions:
- Intraspinal transplantation of BM-iMG cells shows significant therapeutic benefits in an ALS mouse model.
- BM-iMG cells represent a promising cell-based therapy for amyotrophic lateral sclerosis.
- Further research and clinical translation are warranted for applying BM-iMG cell therapy in human ALS patients.
