Human pluripotent stem cell-derived ectomesenchymal stromal cells promote more robust functional recovery than umbilical cord-derived mesenchymal stromal cells after hypoxic-ischaemic brain damage

Abstract

Aims: Hypoxic-ischaemic encephalopathy (HIE) is one of the most serious complications in neonates and infants. Mesenchymal stromal cell (MSC)-based therapy is emerging as a promising treatment avenue for HIE. However, despite its enormous potential, the clinical application of MSCs is limited by cell heterogeneity, low isolation efficiency and unpredictable effectiveness. In this study, we examined the therapeutic effects and underlying mechanisms of human pluripotent stem cell-derived ectomesenchymal stromal cells (hPSC-EMSCs) in a rat model of HIE. Methods: hPSC-EMSCs were induced from either human embryonic stem cells or induced pluripotent stem cells. Stem cells or the conditioned medium (CM) derived from stem cells were delivered intracranially or intranasally to neonatal rats with HIE. Human umbilical cord-derived MSCs (hUC-MSCs) were used as the therapeutic comparison control and phosphate-buffered saline (PBS) was used as a negative control. Lesion size, apoptosis, neurogenesis, astrogliosis and microgliosis were evaluated. The rotarod test and Morris water maze were used to determine brain functional recovery. The PC-12 cell line, rat primary cortical neurons and neural progenitor cells were used to evaluate neurite outgrowth and the neuroprotective and neurogenesis effects of hPSC-EMSCs/hUC-MSCs. RNA-seq and enzyme-linked immunosorbent assays were used to determine the secretory factors that were differentially expressed between hPSC-EMSCs and hUC-MSCs. The activation and suppression of extracellular signal-regulated kinase (ERK) and cAMP response element-binding protein (CREB) were characterised using western blotting and immunofluorescent staining. Results: hPSC-EMSCs showed a higher neuroprotective potential than hUC-MSCs, as demonstrated by a more significant reduction in lesion size and apoptosis in the rat brain following hypoxia-ischaemia (HI). Compared with PBS treatment, hPSC-EMSCs promoted endogenous neurogenesis and alleviated astrogliosis and microgliosis. hPSC-EMSCs were more effective than hUC-MSCs. hPSC-EMSCs achieved a greater recovery of brain function than hUC-MSCs and PBS in rats with HIE. CM derived from hPSC-EMSCs had neuroprotective and neurorestorative effects in vitro through anti-apoptotic and neurite outgrowth- and neurogenesis-promoting effects. Direct comparisons between hPSC-EMSCs and hUC-MSCs revealed the significant enrichment of a group of secretory factors in hPSC-EMSCs, including nerve growth factor (NGF), platelet-derived growth factor-AA and transforming growth factor-β 2 , which are involved in neurogenesis, synaptic transmission and neurotransmitter transport, respectively. Mechanistically, the CM derived from hPSC-EMSCs was found to potentiate NGF-induced neurite outgrowth and the neuronal differentiation of NPCs via the ERK/CREB pathway. Suppression of ERK or CREB abolished CM-potentiated neuritogenesis and neuronal differentiation. Finally, intranasal ...

Mesenchymal Stem Cells: Therapeutic Mechanisms for Stroke

“…In addition, the overexpression of the BDNF gene could direct the MSC into the neural differentiation pathway [ 7 ]. In other aspects, the function of NGF was to prevent neuron apoptosis and to increase neuron proliferation [ 24 ], while VEGF enhanced angiogenesis [ 25 ], and PDGF promoted cell migration, the growth of primary cortical neurons, angiogenesis and axon growth, as well as the inhibition of neuroinflammation [ 26 , 27 , 28 ].…”

Single-cell RNA sequencing uncovers a neuron-like macrophage subset associated with cancer pain

“…LLC conditioned medium (LLC-CM) was collected by overnight incubation of LLC cells (70 to 80% confluence) with serum-free DMEM/F12, followed by filtration with 0.2-μm nylon membrane. The neuronal model PC-12 cells were cultured in RPMI 1640 medium supplemented with 10% horse serum, 5% fetal bovine serum (FBS), and penicillin-streptomycin (100 U/ml) ( 92 ).…”

“…MNTs were induced by further stimulating BMDM with TGF-β1 (5 ng/ml) for 5 days following 10% (v/v) LLC-CM incubation for 7 days. Mouse primary neuron was prepared according to previously established protocol ( 92 ). In brief, cortices were dissected and dissociated by 0.25% trypsin at 37°C for 20 min.…”

Heterogeneity of In Vitro Expanded Mesenchymal Stromal Cells and Strategies to Improve Their Therapeutic Actions

“…Recent studies have shown beneficial effects of iPSC-MSC and their extracellular vesicles for treating different pathologies, including brain diseases [ 158 , 159 ], respiratory diseases with inflammatory symptoms [ 160 , 161 ], vascular-related pathologies [ 162 , 163 ], immune-mediated conditions [ 164 , 165 ] or cancer [ 166 , 167 ]. Using a hypoxic–ischemic rat model, J. Huang and colleagues demonstrated that the intracranial delivery of iPSC-MSCs reduces the brain pathology and restores motor and cognitive function [ 158 ]. In addition, they demonstrated that iPSC-MSCs promote more robust neuroprotective effects than umbilical cord-derived MSCs after brain damage.…”