Stage-specific changes in neurogenic and glial markers in Alzheimer's disease.

Background: Reports of altered endogenous neurogenesis in people with Alzheimer's disease (AD) and transgenic AD models have suggested that endogenous neurogenesis may be an important treatment target, but there is considerable discrepancy among studies. We examined endogenous neurogenesis and glia changes across the range of pathologic severity of AD in people with and without dementia to address this key question.

Methods: Endogenous neurogenesis and glia in the subventricular zone and dentate gyrus neurogenic niches were evaluated using single and double immunohistochemistry and a validated antibody selection for stage-specific and type-specific markers in autopsy tissue from a representative cohort of 28 participants in the Medical Research Council Cognitive Function and Ageing Study.

Developing technologies to unlock the therapeutic and research potential of human stem cells.

Since human embryonic stem cells (hESCs) were first isolated and cultured nearly 15 years ago, stem cell biology has been a promising and fast-moving area of research. Improved clinical predictivity in drug development, use in assays to personalise medicine effectively and as the foundation for cell-based therapies are all areas where stem cells can play an important role. But with opportunities come challenges and it is vital that the field of stem cells continues to progress to achieve its potential.

Screening ethnically diverse human embryonic stem cells identifies a chromosome 20 minimal amplicon conferring growth advantage.

The International Stem Cell Initiative analyzed 125 human embryonic stem (ES) cell lines and 11 induced pluripotent stem (iPS) cell lines, from 38 laboratories worldwide, for genetic changes occurring during culture. Most lines were analyzed at an early and late passage. Single-nucleotide polymorphism (SNP) analysis revealed that they included representatives of most major ethnic groups.

The effects of culture on genomic imprinting profiles in human embryonic and fetal mesenchymal stem cells.

Human embryonic stem (hES) cells and fetal mesenchymal stem cells (fMSC) offer great potential for regenerative therapy strategies. It is therefore important to characterise the properties of these cells in vitro. One major way the environment impacts on cellular physiology is through changes to epigenetic mechanisms.

Increased neural progenitors in vascular dementia.

Since groundbreaking studies demonstrated the presence of progenitor cells in the adult human brain, there have been intense interests in their potential therapeutic application, but to date only limited data has been obtained in man. An immunohistological study was performed in order to examine neurogenesis in both the subventricular and peri-infarct zones of vascular dementia patients compared to age-matched controls. The results were striking, showing a significant increase of progenitor cells in both the subventricular zone and in peri-infarct area in patients with vascular dementia compared to controls, which was sustained even in patients with infarcts occurring more than three months prior to autopsy.

Neural progenitors promote axon growth in vitro and ex vivo but not following injury.

Following an injury to the dorsal roots primary sensory afferents fail to regenerate past the hostile dorsal root entry zone (DREZ), the interface between the peripheral and central nervous system. Neural progenitor cells have previously been utilised as a cellular replacement therapy in a variety of CNS injury models. Here we show for the first time that NPCs are capable of promoting neurite outgrowth from adult sensory neurons in vitro and ex vivo cryo-cultures.

Sequential RARbeta and alpha signalling in vivo can induce adult forebrain neural progenitor cells to differentiate into neurons through Shh and FGF signalling pathways.

We show here the role of retinoic acid receptor (RAR) beta and alpha signalling in proliferation and differentiation of endogenous adult forebrain neural progenitor cells (NPCs). RARbeta activation stimulates Sonic hedgehog signalling (Shh), and induces the proliferation of the NPCs. They can be induced to become Doublecortin (DCX) expressing migrating neuroblasts by RARalpha signalling, some of which differentiate into cholinergic neurons.

Generation of hepatocytes from human embryonic stem cells.

Use of human hepatocytes for therapeutic and drug discovery applications is hampered by limited tissue source and the inability of hepatocytes to proliferate and maintain function long-term in vitro. Human embryonic stem (hES) cells are immortal and pluripotent and may provide a cell source for functional human hepatocytes (1) Here we have outlined some of the protocols currently in use for the generation of hepatocytes from hES cells.

Isolation, in vitro cultivation and characterisation of foetal liver cells.

Hepatocyte transplantation has recently become an efficient clinical method in the treatment of patients with metabolic liver diseases. The shortage of donor cells remains an obstacle to treat more patients. Foetal liver tissues may therefore be useful as an alternative source of generating functional hepatocytes after in vitro culture and maturation.

Homogeneous monocytes and macrophages from human embryonic stem cells following coculture-free differentiation in M-CSF and IL-3.

Objective: To develop a simple and efficient method for producing homogeneous populations of monocytes and macrophages from human embryonic stem cells (hES).

Materials And Methods: Human embryonic stem cell lines KCL001, KCL002, and HUES-2 were differentiated into monocytes by coculture-free differentiation with two growth factors using a three-step method. The method involved embryoid body (EB) formation in hES media, directed differentiation with macrophage colony-stimulating factor and interleukin (IL)-3, and harvest of nonadherent monocytes from the culture supernatants.

Transplanted neural progenitor cells survive and differentiate but achieve limited functional recovery in the lesioned adult rat spinal cord.

Unlabelled: Endogenous repair after injury in the adult CNS is limited by a number of factors including cellular loss, inflammation, cavitation and glial scarring. Spinal cord neural progenitor cells (SCNPCs) may provide a valuable cellular source for promoting repair following spinal cord injury. SCNPCs are multipotent, can be expanded in vitro, have the capacity to differentiate into CNS cell lineages and are capable of long-term survival following transplantation.

Embryonic stem cells: protecting pluripotency from alloreactivity.

There can be little doubt that 2006 turned out to be the annus horribilis for therapeutic cloning by somatic nuclear transfer (SNT). As the full extent of the fraud surrounding the generation of patient-specific embryonic stem (ES) cell lines became apparent, hopes began to fade for the advent of cell replacement therapies (CRT), free from the confounding issues of immune rejection. While the dust begins to settle, it is perhaps pertinent to ask whether the promise of SNT is still worth pursuing or whether alternative strategies for immune evasion might help fill the void.

Characterization of human embryonic stem cell lines by the International Stem Cell Initiative.

The International Stem Cell Initiative characterized 59 human embryonic stem cell lines from 17 laboratories worldwide. Despite diverse genotypes and different techniques used for derivation and maintenance, all lines exhibited similar expression patterns for several markers of human embryonic stem cells. They expressed the glycolipid antigens SSEA3 and SSEA4, the keratan sulfate antigens TRA-1-60, TRA-1-81, GCTM2 and GCT343, and the protein antigens CD9, Thy1 (also known as CD90), tissue-nonspecific alkaline phosphatase and class 1 HLA, as well as the strongly developmentally regulated genes NANOG, POU5F1 (formerly known as OCT4), TDGF1, DNMT3B, GABRB3 and GDF3.

Endogenous neurogenesis in the human brain following cerebral infarction.

Increased endogenous neurogenesis has a significant regenerative role in many experimental models of cerebrovascular diseases, but there have been very few studies in humans. We therefore examined whether there was evidence of altered endogenous neurogenesis in an 84-year-old patient who suffered a cerebrovascular accident 1 week prior to death. Using antibodies that specifically label neural stem/neural progenitor cells, we examined the presence of immunopositive cells around and distant from the infarcted area, and compared this with a control, age-matched individual.

Altered neurogenesis in Alzheimer's disease.

Background: Exciting preliminary work indicates an increase in progenitor activity in the subgranular zone of the dentate gyrus of people with Alzheimer's disease (AD) compared to that of controls. We examine progenitor activity in the other main progenitor niche, the subventricular zone (SVZ), as well as potential associations with key pathological and neurochemical substrates.

Method: Immunocytochemistry techniques utilizing nestin and Musashi1 antibodies were used to examine progenitor activity in the SVZ and to enable comparisons between seven patients with AD and seven controls, based upon the quantification of the percentage area covered, using the Image Pro Plus v.

Selective effects of the APOE epsilon4 allele on presynaptic cholinergic markers in the neocortex of Alzheimer's disease.

The effects of the APOE epsilon4 allele on a range of pre- and postsynaptic cholinergic markers were studied in a cohort of community-based Alzheimer's disease (AD) patients. Compared with age-matched controls, the postmortem AD neocortex showed decreased choline acetyltransferase (ChAT) and acetyl cholinesterase activities, lower muscarinic M2, and nicotinic alpha4beta2 receptor densities, as well as reduced M1 receptor coupling to G-proteins. However, the epsilon4 allele was dose-dependently correlated only with higher losses of ChAT activities.

Derivation and characterization of four new human embryonic stem cell lines: the Danish experience.

In September 2003, legislation approved in Denmark legalized work on surplus human embryos from IVF for clinical purposes to establish human embryonic stem (ES) cell cultures. The aim of this study was to establish such stem cell lines. Fresh surplus embryos were donated after informed consent from the donors.

Regenerative medicine in Parkinson's disease: generation of mesencephalic dopaminergic cells from embryonic stem cells.

Cell replacement therapy has been proposed as a means of replacing specific populations of cells lost through trauma, disease or ageing. Parkinson's disease is a progressive neurodegenerative disorder caused by the loss of midbrain dopaminergic neurons. Intrastriatal transplants of human foetal mesencephalic tissue in Parkinson's patients have demonstrated clinical efficacy, but the limited availability of tissue precludes systematic use of this treatment.

Generation of a human embryonic stem cell line encoding the cystic fibrosis mutation deltaF508, using preimplantation genetic diagnosis.

Human embryonic stem (hES) cells are pluripotent cells isolated from early human embryos. They can be grown in vitro and made to differentiate into many different cell types. These properties have suggested that they may be useful in cell replacement therapy for many degenerative diseases.

Generation of insulin-expressing cells from mouse embryonic stem cells.

The therapeutic potential of transplantation of insulin-secreting pancreatic beta-cells has stimulated interest in using pluripotent embryonic stem (ES) cells as a starting material from which to generate insulin secreting cells in vitro. Mature beta-cells are endodermal in origin so most reported differentiation protocols rely on the identification of endoderm-specific markers. However, endoderm development is an early event in embryogenesis that produces cells destined for the gut and associated organs in the embryo, and for the development of extra-embryonic structures such as the yolk sac.

Timing of the retinoid-signalling pathway determines the expression of neuronal markers in neural progenitor cells.

By culturing neural progenitor cells in the presence of retinoid receptor agonists, we have defined the components of the retinoid-signalling pathway that are important for the birth and maintenance of neuronal cells. We provide evidence that depending on the order and combination of retinoid receptors activated, different neuronal cells are obtained. Astrocytes and oligodendrocytes are predominantly formed in the presence of activated retinoic acid receptor (RAR) alpha, whereas motoneurons are formed when RARbeta is activated.

The in vitro differentiation of rat neural stem cells into an insulin-expressing phenotype.

Mature beta-cells and nerve cells share many functional similarities despite originating from different embryonic germ layers. The aim of this study was to investigate the potential of neural stem cells (NSCs), isolated from foetal rat brain, as a starting material from which to generate functionally responsive, insulin-containing cells. Our results demonstrated that NSCs can be significantly expanded in vitro and can be induced to express increased preproinsulin mRNA levels.