Neurotrophic Factor Secretion and Neural Differentiation Potential of Multilineage-differentiating Stress-enduring (Muse) Cells Derived from Mouse Adipose Tissue

Abstract

Multilineage-differentiating stress-enduring (Muse) cells are endogenous pluripotent stem cells that can be isolated based on stage-specific embryonic antigen-3 (SSEA-3), a pluripotent stem cell-surface marker. However, their capacities for survival, neurotrophic factor secretion, and neuronal and glial differentiation are unclear in rodents. Here we analyzed mouse adipose tissue-derived Muse cells in vitro. We collected mesenchymal stem cells (MSCs) from C57BL/6 J mouse adipose tissue and separated SSEA-3+, namely Muse cells, and SSEA-3–, non-Muse cells, to assess self-renewability; pluripotency marker expression (Nanog, Oct3/4, Sox2, and SSEA-3); spontaneous differentiation into endodermal, mesodermal, and ectodermal lineages; and neural differentiation capabilities under cytokine induction. Neurally differentiated Muse and non-Muse cell functions were assessed by calcium imaging. Antioxidant ability was measured to assess survival under oxidative stress. Brain-derived neurotrophic factor (BDNF), vascular endothelial cell growth factor (VEGF), and hepatocyte growth factor (HGF) secretion were analyzed in enzyme-linked immunosorbent assays. SSEA-3+ Muse cells (6.3 ± 1.9% of mouse adipose-MSCs), but not non-Muse cells, exhibited self-renewability, spontaneous differentiation into the three germ layers, and differentiation into cells positive for Tuj-1 (27 ± 0.9%), O4 (17 ± 3.4%), or GFAP (23 ± 1.3%) under cytokine induction. Neurally differentiated Muse cells responded to KCl depolarization with greater increases in cytoplasmic Ca2+ levels than non-Muse cells. Cell survival under oxidative stress was significantly higher in Muse cells (50 ± 2.7%) versus non-Muse cells (22 ± 2.8%). Muse cells secreted significantly more BDNF, VEGF, and HGF (273 ± 12, 1479 ± 7.5, and 6591 ± 1216 pg/mL, respectively) than non-Muse cells (133 ± 4.0, 1165 ± 20, and 2383 ± 540 pg/mL, respectively). Mouse Muse cells were isolated and characterized for the first time. Muse cells showed greater pluripotency-like characteristics, survival, neurotrophic factor secretion, and neuronal and glial-differentiation capacities than non-Muse cells, indicating that they may have better neural-regeneration potential.

Therapeutic benefit of Muse cells in a mouse model of amyotrophic lateral sclerosis

“…Reactive astrocytes are an important therapeutic target of ALS 29 , and the microenvironmental signal of the ALS mice spinal cord may differentiate Muse cells into GFAP-positive astrocytes, especially into A2 astrocytes, which might secrete molecules that provide neurotrophic support and modulate inflammatory responses 30 . Muse cells themselves can produce various neurotrophic factors, including brain-derived neurotrophic factor (BDNF), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), insulin-like growth factor 1 (IGF-1), epidermal growth factor (EGF), prostaglandin E2 (PGE2), and angiopietin-1 (Ang1) 16 – 19 , 31 . Therefore, they might have supplied beneficial factors to motor neurons and astrocytes, preventing myofiber atrophy in the ALS model.…”

Verotoxin Receptor-Based Pathology and Therapies

“…Muse cells home to sites of tissue damage and there differentiate to regenerate the damaged tissue type (Young, 2018). This includes neuronal cells (Nitobe et al, 2019). There are many current clinical trials based on these cells (Dezawa, 2018;Kuroda et al, 2018).…”

Molecular Profiles of Cell-to-Cell Variation in the Regenerative Potential of Mesenchymal Stromal Cells

“…SSEA-3 + CD105 + cells account for as little as 1% of hBM-MSCs and 3-9% of hAD-MSCs [74, 75]. MUSE cells express pluripotency factors, such as octamer-binding transcription factor 3/4 (OCT3/4) and SOX2 [74, 76], and exhibit triploblastic differentiation at the single-cell level [77]. This pluripotent phenotype is characteristic of MUSE cells isolated from various tissues, including human bone marrow, skin, and adipose [74, 77, 78].…”