Latent Tri-lineage Potential of Human Menstrual Blood–Derived Mesenchymal Stromal Cells Revealed by Specific In Vitro Culture Conditions

Abstract

Human menstrual blood-derived mesenchymal stromal cells (MenSCs) have become not only an important source of stromal cells for cell therapy but also a cellular source for neurologic disorders in vitro modeling. By using culture protocols originally developed in our laboratory, we show that MenSCs can be converted into oating neurospheres (NSs) using the Fast-N-Spheres medium for 24-72h, and can be transdifferentiated into functional dopaminergic-like (DALNs, ~26% TH+/DAT+ ow cytometry) and cholinergic-like neurons (ChLNs, ~46% ChAT+/VAChT ow cytometry) which responded to dopamine-and acetylcholine-triggered neuronal Ca 2+ inward stimuli when cultured with the NeuroForsk and the Cholinergic-N-Run medium , respectively in a timely fashion (i.e., 4-7 days). Here, we also report a direct transdifferentiation method to induce MenSCs into functional astrocyte-like cells (ALCs) by incubation of MenSCs in commercial Gibco® Astrocyte Medium in 7-days. The MSCs-derived ALCs (~59% GFAP+/S100b+) were found to respond to glutamate-induced Ca 2+ inward stimuli. Altogether these results show that MenSCs are a reliable source to obtain functional neurogenic cells to further investigate the neurobiology of neurologic disorders.

Cannabidiol Protects Dopaminergic-like Neurons against Paraquat- and Maneb-Induced Cell Death through Safeguarding DJ-1CYS106 and Caspase 3 Independently of Cannabinoid Receptors: Relevance in Parkinson’s Disease

“…Previous observations have demonstrated that MenSCs cultured in NeuroForsk medium can transdifferentiate into functional dopaminergic-like neurons (DALNs) according to the dopaminergic markers DAT (33%) and TH (29%). Accordingly, we confirmed that MenSCs can transdifferentiate into DALNs and the percentage of cellular transdifferentiation markers were similar (DAT 43%, TH 32%) as assessed by the flow cytometry assay (Figure A).…”

(−)-Epigallocatechin-3-Gallate Diminishes Intra-and Extracellular Amyloid-Induced Cytotoxic Effects on Cholinergic-like Neurons from Familial Alzheimer’s Disease PSEN1 E280A

“…Using ALCs as a cellular model of human brain astrocytes [ 29 ], we demonstrate for the first time that the supernatant culture medium from PSEN1 E280A ChLNs, but not the supernatant culture medium from WT ChLNs, induces the activation of NF-κB—a proinflammatory transcription factor [ 57 ] and the secretion of pro-inflammatory cytokine IL-6 [ 26 ] in WT MenSCs-derived astrocyte cells. Here, we also report for the first time that EGCG—a well-established anti-inflammatory molecule (e.g., [ 22 ]) dramatically reduced the activation of NF-κB and the secretion of pro-inflammatory cytokine in ALCs.…”

“…Isolation of mesenchymal stromal cells (MenSCs) derived from wild-type human menstrual blood were obtained from tissue bank code (TBC# 69308)-UdeA [ 29 ]. For astrocyte differentiation, 1 × 10 4 WT MenSCs/cm 2 were seeded in 25 cm 2 culture flasks in regular culture medium (RCm, DMEM low glucose Sigma cat# D6046 media supplemented with 10% FBS) until reach 40% of confluence.…”

“…Due to the importance of astrocytes in brain homeostasis and neuroinflammation, they have become the focus of active research in AD [ 27 , 28 ]. Recently, we have shown that functional astrocyte-like cells (ALCs, ~59% GFAP+/S100β+ responsive to glutamate-induced Ca 2+ inward stimuli) can be derived from menstrual stromal cells (MenSCs) by direct transdifferentiation method using commercial Gibco ® Astrocyte medium for 7 days [ 29 ]. Interestingly, it has been shown that Aβ42 can trigger reactive astrogliosis, releasing neuroinflammatory cytokines (e.g., IL-6) and via activation of transcription factor NF-κB [ 30 ].…”

Polycationic peptide R7-G-Aβ25-35 selectively induces cell death in leukemia Jurkat T cells through speedy mitochondrial depolarization, and CASPASE-3 -independent mechanism

“…Isolation of MenSCs was performed according to previous reports [ 31 ]. Briefly, a menstrual blood sample ( t issue b ank c ode (TBC # 69308) was delivered to the laboratory and mixed with an equal volume of phosphate buffer saline (PBS) containing 1 mM ethylenediamine tetra-acetic acid (EDTA), with 100 U/mL penicillin/streptomycin 0.25 mg/mL amphotericin B, and standard Ficoll procedures.…”