Rapid and efficient generation of neural progenitors from adult bone marrow stromal cells by hypoxic preconditioning

Abstract

BackgroundBone marrow stromal cells (BMSCs) are attractive as a source of neural progenitors for ex vivo generation of neurons and glia. Limited numbers of this subpopulation, however, hinder translation into autologous cell-based therapy. Here, we demonstrate rapid and efficient conditioning with hypoxia to enrich for these neural progenitor cells prior to further expansion in neurosphere culture.MethodAdherent cultures of BMSCs (rat/human) were subjected to 1 % oxygen for 24 h and then subcultured as neurospheres with epidermal growth factor (EGF) and basic fibroblast growth factor supplementation. Neurospheres and cell progeny were monitored immunocytochemically for marker expression. To generate Schwann cell-like cells, neurospheres were plated out and exposed to gliogenic medium. The resulting cells were co-cultured with purified dorsal root ganglia (rat) neurons and then tested for commitment to the Schwann cell fate. Fate-committed Schwann cells were subjected to in vitro myelination assay.ResultsTransient hypoxic treatment increased the size and number of neurospheres generated from both rat and human BMSCs. This effect was EGF-dependent and attenuated with the EGF receptor inhibitor erlotinib. Hypoxia did not affect the capacity of neurospheres to generate neuron- or glia-like precursors. Human Schwann cell-like cells generated from hypoxia-treated BMSCs demonstrated expression of S100β /p75 and capacity for myelination in vitro.ConclusionEnhancing the yield of neural progenitor cells with hypoxic preconditioning of BMSCs in vitro but without inherent risks of genetic manipulation provides a platform for upscaling production of neural cell derivatives for clinical application in cell-based therapy.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-016-0409-x) contains supplementary material, which is available to authorized users.

The Role of Hypoxia on the Neuronal Differentiation of Gingival Mesenchymal Stem Cells: A Transcriptional Study

“…However, hypoxia did not alter the ability of neurospheres to form neuron and glia precursors. In addition, Schwann celllike cells obtained from hypoxia-preconditioned BMSCs expressed S100b /p75 and the ability for myelination in vitro 44 .…”

The effect of hypoxia preconditioning on the neural and stemness genes expression profiling in human umbilical cord blood mesenchymal stem cells

“…In another study that conducted with KL Mung and colleagues, demonstrated that bone marrow mesenchymal stem cells express GFAP under hypoxia condition [30]. These proteins produce a network for cell protection and strength.…”

Human dental stem cell derived transgene‐free iPSCs generate functional neurons via embryoid body‐mediated and direct induction methods

“…Using mesenchymal stromal/stem cells (MSCs) as a source for neurogenesis has been tested rigorously in the hope that non‐NSCs may be used as a cell source for neural regeneration. However, the results have been controversial and inconsistent (Aanismaa, Hautala, Vuorinen, Miettinen, & Narkilahti, ; Arthur, Rychkov, Shi, Koblar, & Gronthos, ; Chen et al, ; Deng, Petersen, Steindler, Jorgensen, & Laywell, ; Gervois et al, ; Jung, Kim, Moon, Hong, & Hyun, ; Mung et al, ; Pittenger et al, ; Shiota et al, ; Tondreau et al, ; Wehner et al, ). Many reports attempted to define NSC properties of MSCs only by detecting gene expression but not examining or demonstrating the functional electrophysiology that is the critical characteristic of neurons.…”