ABSTRACT
Malignant gliomas contain a population of self-renewing tumorigenic stem-like cells; however, it remains unclear how these glioma stem cells (GSCs) self-renew or generate cellular diversity at the single-cell level. Asymmetric cell division is a proposed mechanism to maintain cancer stem cells, yet the modes of cell division that GSCs utilize remain undetermined. Here, we used single-cell analyses to evaluate the cell division behavior of GSCs. Lineage-tracing analysis revealed that the majority of GSCs were generated through expansive symmetric cell division and not through asymmetric cell division. The majority of differentiated progeny was generated through symmetric pro-commitment divisions under expansion conditions and in the absence of growth factors, occurred mainly through asymmetric cell divisions. Mitotic pair analysis detected asymmetric CD133 segregation and not any other GSC marker in a fraction of mitoses, some of which were associated with Numb asymmetry. Under growth factor withdrawal conditions, the proportion of asymmetric CD133 divisions increased, congruent with the increase in asymmetric cell divisions observed in the lineage-tracing studies. Using single-cell-based observation, we provide definitive evidence that GSCs are capable of different modes of cell division and that the generation of cellular diversity occurs mainly through symmetric cell division, not through asymmetric cell division.
Subject(s)
Antigens, CD/metabolism , Glioma/pathology , Glycoproteins/metabolism , Neoplastic Stem Cells/metabolism , Peptides/metabolism , AC133 Antigen , Antigens, CD/analysis , Cell Division , Cell Lineage , Epidermal Growth Factor/pharmacology , Fibroblast Growth Factor 2/pharmacology , Glioma/metabolism , Glycoproteins/analysis , Humans , Laminin/metabolism , Mitosis , Neoplastic Stem Cells/pathology , Peptides/analysisABSTRACT
The identification and characterization of multipotent neural precursors open the possibility of transplant therapies, but this approach is complicated by the widespread pathology of many degenerative diseases. Activation of endogenous precursors that support regenerative mechanisms is a possible alternative. We have previously shown that Notch ligands promote stem cell survival in vitro. Here, we show that there is an intimate interaction between insulin and Notch receptor signaling. Notch ligands also expand stem cell numbers in vivo with correlated benefits in brain ischemia. We now show that insulin promotes recovery of injured dopamine neurons in the adult brain. This response suggests that activating survival mechanisms in neural stem cells will promote recovery from progressive degenerative disease.
Subject(s)
Brain Diseases/metabolism , Neurons/metabolism , Stem Cells/metabolism , Adult , Animals , Basic Helix-Loop-Helix Transcription Factors/metabolism , Brain Diseases/drug therapy , Brain Diseases/pathology , Cell Survival/drug effects , Dopamine/metabolism , Humans , Insulin/metabolism , Insulin/pharmacology , Intracellular Signaling Peptides and Proteins , Ligands , Membrane Proteins/metabolism , Membrane Proteins/pharmacology , Nerve Tissue Proteins/metabolism , Neurons/drug effects , Neurons/pathology , Oxidopamine/toxicity , Rabbits , Rats , Receptors, Notch/metabolism , Repressor Proteins , Signal Transduction , Stem Cells/drug effects , Stem Cells/pathologyABSTRACT
The use of cell-based therapy may be a valid therapeutic approach to ischemic brain injury. Stem cells have been proposed as a new form of cell based therapy in a variety of disorders, including acute and degenerative brain diseases. Up to date most efforts have concentrated on transplantation of embryonic stem cells (ESC) or neural stem cells (NSCs) obtained from immortalized cell lines into the diseased brain. These procedures require harvesting the appropriate stem cell, expansion in vitro and transplantation. Endogenous NSCs have been identified in the central nervous system where they reside largely in the subventricular zone and in the subgranular zone of the hippocampus. Endogenous NSCs may be capable of self-renewal and differentiation into functional glia and neurons. Manipulation of endogenous NSCs may bypass the need to use ESC as a form of therapy thus avoiding the complex ethical and biological issues involved with ES cells or immortalized cell lines. This review summarizes the evidence recently gathered in support of a therapeutic role for endogenous NSCs in acute experimental stroke.
Subject(s)
Brain Ischemia/therapy , Neurons/physiology , Recovery of Function/physiology , Stem Cells/physiology , Animals , Cell Survival , Humans , Models, Neurological , Stem Cell Transplantation/methodsABSTRACT
Embryonic stem (ES) cells have revolutionised our understanding of animal physiology. Analysis of chimaeric mice generated from these cells allows us to study the role of genes in development and function of the nervous system. The NMDA receptor, one of the two major ionotropic glutamate receptors, has been proposed to play fundamental roles in the survival, migration, differentiation, and activity-dependent maturation of neural cells. The NMDA receptor subunit 1 (NR1) gene is indispensable for receptor function, and knock-out mice die at birth, inhibiting the study of glutamate signalling in postnatal neurons. Homozygous NR1-/- ES cells were derived from matings of heterozygous mice under feeder-free conditions. Chimaeras were made by incorporating these ES cells into wild-type blastocysts and by the classical aggregation of morulae between wild-type and NR1-/- embryos. The resulting chimaeras survive and develop normally. NR1-/- neurons, identified by their lacZ label, were analysed and quantified in developing and adult brains with varying knock-out contributions in every single brain region. Specifically, postnatal ontogenesis of cerebellum and hippocampus was normal. Accordingly, in chimaeric mice, NMDA receptor-initiated signals are not required for the migration, differentiation, and survival of most types of neurons in the central nervous system, in a cell-autonomous way.