Neurogenic and neuro-protective potential of a novel subpopulation of peripheral blood-derived CD133+ ABCG2+CXCR4+ mesenchymal stem cells: development of autologous cell-based therapeutics for traumatic brain injury

Abstract

IntroductionNervous system injuries comprise a diverse group of disorders that include traumatic brain injury (TBI). The potential of mesenchymal stem cells (MSCs) to differentiate into neural cell types has aroused hope for the possible development of autologous therapies for central nervous system injury.MethodsIn this study we isolated and characterized a human peripheral blood derived (HPBD) MSC population which we examined for neural lineage potential and ability to migrate in vitro and in vivo. HPBD CD133+, ATP-binding cassette sub-family G member 2 (ABCG2)+, C-X-C chemokine receptor type 4 (CXCR4)+ MSCs were differentiated after priming with β-mercaptoethanol (β-ME) combined with trans-retinoic acid (RA) and culture in neural basal media containing basic fibroblast growth factor (FGF2) and epidermal growth factor (EGF) or co-culture with neuronal cell lines. Differentiation efficiencies in vitro were determined using flow cytometry or fluorescent microscopy of cytospins made of FACS sorted positive cells after staining for markers of immature or mature neuronal lineages. RA-primed CD133+ABCG2+CXCR4+ human MSCs were transplanted into the lateral ventricle of male Sprague-Dawley rats, 24 hours after sham or traumatic brain injury (TBI). All animals were evaluated for spatial memory performance using the Morris Water Maze (MWM) Test. Histological examination of sham or TBI brains was done to evaluate MSC survival, migration and differentiation into neural lineages. We also examined induction of apoptosis at the injury site and production of MSC neuroprotective factors.ResultsCD133+ABCG2+CXCR4+ MSCs consistently expressed markers of neural lineage induction and were positive for nestin, microtubule associated protein-1β (MAP-1β), tyrosine hydroxylase (TH), neuron specific nuclear protein (NEUN) or type III beta-tubulin (Tuj1). Animals in the primed MSC treatment group exhibited MWM latency results similar to the uninjured (sham) group with both groups showing improvements in latency. Histological examination of brains of these animals showed that in uninjured animals the majority of MSCs were found in the lateral ventricle, the site of transplantation, while in TBI rats MSCs were consistently found in locations near the injury site. We found that levels of apoptosis were less in MSC treated rats and that MSCs could be shown to produce neurotropic factors as early as 2 days following transplantation of cells. In TBI rats, at 1 and 3 months post transplantation cells were generated which expressed markers of neural lineages including immature as well as mature neurons.ConclusionsThese results suggest that PBD CD133+ABCG2+CXCR4+ MSCs have the potential for development as an autologous treatment for TBI and neurodegenerative disorders and that MSC derived cell products produced immediately after transplantation may aid in reducing the immediate cognitive defects of TBI.

Cell-based Therapy for Acute Organ Injury

“…The underlying mechanisms were mainly attributed to a microvesicle induced up-regulation of anti-apoptotic genes (Bcl-xL, Bcl2) and to a down-regulation of apoptotic genes (caspase-1, caspase-8, lymphotoxin-α) in tubular epithelial cells. A similar decrease in apoptotic genes expression (caspase-3 pathway) and up-regulation of phosphorylated protein kinase B pro-survival pathway leading to new neuron generation 139,140 were found in TBI treated with MSC 136,141 . Finally, the over expression of genes involved in the anti-apoptotic pathways (such as growth hormone and insulin growth factor-1 signaling) also played a therapeutic role in a model of sepsis treated with MSC 78,79 (Figure 5).…”

“…Most preclinical studies in TBI have used BM-MSC 39,136,137,139,252–255 , except two studies which used peripheral blood-derived 141 or umbilical cord-derived MSC 91 . Rats were the most frequent small animal used 39,137,139,141,252–256 , although, a few studies with TBI have been performed in mice 91 .…”

“…Most preclinical studies in TBI have used BM-MSC 39,136,137,139,252–255 , except two studies which used peripheral blood-derived 141 or umbilical cord-derived MSC 91 . Rats were the most frequent small animal used 39,137,139,141,252–256 , although, a few studies with TBI have been performed in mice 91 . In these studies, MSC were typically given from 24 hours to 7 days following TBI and, doses varied from 6 × 10 6 91 to 3.2 × 10 8 254 cells/kg depending on the administration route, which included intravenous 39,136,137,139,140,252–255,257 or intracerebral 91,140,141 .…”

Treating childhood traumatic brain injury with autologous stem cell therapy

“…Acute treatment and injection are imperative for decreasing inflammation as well as ameliorating the secondary cell death cascade. By this attribute, acute injection displays immediate therapeutic effects as well as lessening the need for potential chronic treatment [3, 40]. There is great potential for using acute and chronic injections in tandem through the use of booster shots to further attenuate the multi-faceted benefit of stem cells on brain injury.…”

Differentiation of adult rat mesenchymal stem cells to GABAergic, dopaminergic and cholinergic neurons

“…A small number 33 of MSCs express the CXCR4 (chemokine receptor type 4) receptor 34 and as a result can migrate into SDF-1 (stromal derived factor-1) 35 niches. MSCs with a migration capability seem to be the best Pharmacological Reports xxx (2014) xxx-xxx 36 population to investigate because after transplantation they can 37 migrate to precisely the damaged region [6]. 38 Several …”