Abstract
Mesenchymal stem cells (MSCs) can ameliorate symptoms in several neurodegenerative diseases. However, the toxic environment of a degenerating central nervous system (CNS) characterized by hypoxia, glutamate (Glu) excess and amyloid beta (Abeta) pathology may hamper the survival and regenerative/replacing capacities of engrafted stem cells. Indeed, human MSC (hMSC) exposed to hypoxia were disabled in (i) the capacity of their muscarinic receptors (mAChRs) to respond to acetylcholine (ACh) with a transient increase in intracellular [Ca2+], (ii) their capacity to metabolize Glu, reflected by a strong decrease in glutamine synthetase activity, and (iii) their survival on exposure to Glu. Cocultivation of MSC with PC12 cells expressing the amyloid precursor protein gene (APPsw-PC12) increased the release of IL-6 from MSC. HMSC exposed to erythropoietin (EPO) showed a cholinergic neuron-like phenotype reflected by increased cellular levels of choline acetyltransferase, ACh and mAChR. All their functional deficits observed under hypoxia, Glu exposure and APPsw-PC12 cocultivation were reversed by the application of EPO, which increased the expression of Wnt3a. EPO also enhanced the metabolism of Abeta in MSC by increasing their neprilysin content. Our data show that cholinergic neuron-like differentiation of MSC, their functionality and resistance to a neurotoxic environment is regulated and can be improved by EPO, highlighting its potential for optimizing cellular therapies of the CNS.
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Abbreviations
- Abeta:
-
amyloid beta
- ACh:
-
acetylcholine
- APPsw:
-
amyloid precursor protein Swedish mutation
- ChAT:
-
choline acetyltranferase
- CRD Fzd 7:
-
cystein-rich domain of frizzled 7
- DMEM:
-
Dulbecco's modified Eagle's medium
- EPO:
-
erythropoietin
- EPOR:
-
erythropoietin receptor
- EPI:
-
erythropoietin preincubation
- FCS:
-
fetal calf serum
- GLAST:
-
glutamate-aspartate transporter
- Glu:
-
glutamate
- GS:
-
glutamine synthetase
- H:
-
hypoxia
- HCs:
-
hypoxic conditions
- LDH:
-
lactate dehydrogenase
- LIF:
-
leukemia-inhibitory factor
- M1R:
-
muscarinic receptor subtype 1
- M1R-M3R:
-
muscarinic receptors of type 1–3
- MSCs:
-
mesenchymal stem cells
- N:
-
normoxia
- NCs:
-
normoxic conditions
- NeuN:
-
neuronal nuclei-specific protein
- NGF:
-
nerve growth factor
- PBS:
-
phosphate-buffered saline
- ROX:
-
carboxy-X-rhodamine
- TUNEL:
-
terminal deoxynucleotide transferase-mediated dUTP-biotin nick end labeling.
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Acknowledgements
The technical assistance by Mrs. Ursula Hermanutz-Klein is gratefully acknowledged. We appreciate the support of Dr. Martina Toelge (Microbionix, Regensburg, Germany) in performing multiplex analysis.
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Danielyan, L., Schäfer, R., Schulz, A. et al. Survival, neuron-like differentiation and functionality of mesenchymal stem cells in neurotoxic environment: the critical role of erythropoietin. Cell Death Differ 16, 1599–1614 (2009). https://doi.org/10.1038/cdd.2009.95
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DOI: https://doi.org/10.1038/cdd.2009.95
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